# Dot Blot Technique – Principle, Procedure and Applications

> Dot blot technique detects specific proteins, DNA, or RNA on a membrane. Explore its principle, protocol, membranes, applications, and result interpretation.

Canonical URL: https://biologynotesonline.com/dot-blot/
Author: Sourav Pan
Last updated: October 19, 2026

![Dot Blot Technique – Principle, Procedure and Applications](https://biologynotesonline.com/wp-content/uploads/2024/04/dot-blot-principle-immunodetection-hybridization.webp)

Dot blot technique is a laboratory method used for detection of specific proteins or nucleic acids present in a biological sample. The sample is directly applied on a suitable membrane as small dots, without separation by gel electrophoresis. Sample molecules remain retained on the membrane. The particular target is then detected from these dots.

Dot blotting can be used for both protein and nucleic acid analysis, but the detection method is different. In protein detection, specific antibodies bind with the target protein by antigen-antibody reaction. It is an immunoassay. For deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), a labeled complementary nucleic acid probe is used and binds with the specific sequence by dot blot hybridization.

The bound antibody or probe is detected using a suitable detection system, which produces a signal at the respective dot.

Unlike conventional blotting involving electrophoresis, the molecules are not separated according to their size in dot blot. It generally gives no information about molecular weight of the proteins or length of the nucleic acids.

## What is Dot Blot?

Dot blot is a membrane-based analytical technique used to detect a specific protein or nucleic acid present in a sample. In this technique, the sample is applied directly onto the membrane as a small dot and the particular target is detected from the immobilized sample. The sample is not first separated by [gel electrophoresis](https://biologynotesonline.com/gel-electrophoresis-system/).

For protein analysis, specific antibodies are used which bind with the target protein by antigen-antibody reaction. This antibody-based form of dot blot is referred to as a dot blot immunoassay.

In nucleic acid dot blotting, deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) is immobilized on a membrane. A labeled complementary nucleic acid probe is then used for detection of the specific sequence by hybridization.

Since the sample molecules are not separated before detection, dot blot generally detects the particular target without giving its molecular-size information.

## Purpose of Dot Blot

- To detect a specific protein or antigen in a biological sample by antigen-antibody reaction.

- To identify a particular deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) sequence using a complementary labeled probe.

- To estimate the relative amount of a target protein or nucleic acid present in different samples.

- To screen a large number of samples on a membrane without prior electrophoretic separation.

## Principle of Dot Blot

The principle of dot blot is based on immobilization of protein or nucleic acid sample on a solid membrane and specific detection of the required target. The sample molecules remain bound at the spotted area of the membrane. A specific antibody is used for protein, whereas a complementary nucleic acid probe detects the specific nucleic acid sequence. A signal is then produced at the respective dot where specific binding has occurred.

The membrane can also bind antibody, probe or other detection molecules nonspecifically. This produces background signal. Free binding sites of the membrane are blocked and washing removes the unbound or weakly bound detection molecules. The specificity of dot blot is affected by these conditions. Detection chemistry converts the specifically bound antibody or probe into a visible, fluorescent or chemiluminescent signal.

![Two dot blot detection pathways showing antibodies binding immobilized proteins and complementary probes hybridizing with DNA or RNA on separate membranes.](https://biologynotesonline.com/wp-content/uploads/2024/04/dot-blot-principle-immunodetection-hybridization-1024x725.webp)Two dot blot detection pathways showing antibodies binding immobilized proteins and complementary probes hybridizing with DNA or RNA on separate membranes.

### Principle of Protein Dot Blot Immunoassay

In protein dot blot immunoassay, the protein sample is immobilized directly on the membrane. A specific antibody binds with the respective antigenic site of the target protein by [antigen-antibody reaction](https://biologynotesonline.com/antigen-antibody-interaction/). This antibody-based detection forms the principle of dot blot immunoassay.

The target can be detected directly by a labeled primary antibody. In an indirect detection system, an unlabeled primary antibody first binds with the target protein and a labeled secondary antibody then binds with the primary antibody. The secondary antibody system can increase the detectable signal. Its label produces fluorescence or an enzyme-dependent chemiluminescent signal.

Before antibody detection, the remaining free sites of membrane are blocked with a suitable blocking substance. It reduces nonspecific binding of antibodies with the membrane. Washing is also carried out after antibody incubation to remove unbound or weakly bound antibodies. Poor blocking or insufficient washing can produce high background and false or unclear dot signal.

### Principle of DNA and RNA Dot Blot Hybridization

Dot blot hybridization is based on complementary base pairing between the immobilized nucleic acid and a labeled [nucleic acid probe](https://biologynotesonline.com/hybridization-probes-definition-application/). Deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) is fixed on the membrane. A probe having complementary sequence binds with the particular target sequence by nucleic acid hybridization. The nonhybridized probes are removed by washing.

The target sequence must remain accessible for probe binding. DNA is generally made single-stranded so that the complementary bases become available to the probe.

Temperature, salt concentration and other hybridization conditions affect the binding between probe and target. Washing stringency also controls this binding. More stringent washing removes weakly bound or mismatched probes, whereas unsuitable conditions may increase nonspecific signal or remove the desired hybrid.

The retained probe is detected from its label. Radioactive, enzyme-linked or other detectable probe systems can be used, depending on the assay. Dot blot hybridization is used for detection of a particular DNA or RNA sequence and for comparison of relative target amounts.

Some nucleic acid dot blots use antibodies instead of complementary probes. The antibodies can recognize particular nucleic acid modifications or structures after immobilization on the membrane. It is an antibody-based detection and is different from sequence-specific dot blot hybridization.

## Requirements for Dot Blot

### Membranes Used in Dot Blot

- Nitrocellulose membrane - It is commonly used for binding and immobilization of proteins in protein dot blot.

- Polyvinylidene fluoride (PVDF) membrane - It is another protein-binding membrane used in dot blot. The membrane is prewetted with methanol before use if required by the protocol.

- Nylon membrane - Mainly used for DNA and RNA dot blot. Positively charged nylon membranes are commonly used for the immobilization of nucleic acids.

- Membrane preparation - A suitable membrane is selected depending on the sample and detection method. The membrane is prepared before applying the sample.

### Reagents Required

- Sample - Prepared protein, DNA or RNA sample containing the target molecule.

- Buffers - Different buffers are required for sample preparation, washing and incubation. The type of buffer depends on the assay.

- Blocking solution - It is mainly used in antibody-based detection to block the free binding sites on the membrane. It reduces nonspecific binding of antibodies.

- Antibodies - A specific primary antibody is used for the detection of target proteins. A labeled secondary antibody may also be required.

- Nucleic acid probe - A labeled complementary probe is required for DNA or RNA dot blot. Suitable hybridization reagents are also used.

- Detection reagents - Suitable substrate or imaging reagents are required depending on the type of label used.

### Equipment Required

- [Micropipettes](https://biologynotesonline.com/micropipettes-parts-types-applications-techniques/) and tips - Used for accurate preparation and spotting of samples on the membrane.

- Dot blot manifold - It is used to apply multiple samples at specific positions on the membrane. A compatible manifold can be connected to a vacuum system.

- Sample tubes and trays - Required for sample preparation, blocking, washing and incubation of the membrane with antibodies or probes.

- Incubation equipment - Rocker, shaker or suitable incubator is used depending on the assay conditions.

- Detection system - Chemiluminescence, fluorescence or other suitable imaging systems are used for detecting and recording the dot signals.

## Dot Blot Procedure

### General Dot Blot Workflow

- Prepare the protein, DNA or RNA sample at a suitable concentration. Serial dilutions may be prepared for comparing the signal intensity.

- Select a suitable membrane according to the sample. Nitrocellulose or PVDF membranes are commonly used for proteins, while nylon membranes are used for DNA and RNA.

- Apply the prepared sample directly on the membrane using a micropipette or dot blot manifold. The sample gets immobilized at the respective position in the form of a dot.

- The membrane is then treated according to the target molecule. Proteins are detected using specific antibodies after blocking, whereas DNA and RNA are detected using labeled complementary probes.

- Wash the membrane to remove unbound antibodies or probes. The signals are then detected using a suitable detection system.

### Protein Dot Blot Protocol

![Protein dot blot workflow showing sample spotting on a membrane, blocking, primary and secondary antibody incubation, washing, and detection of circular spots.](https://biologynotesonline.com/wp-content/uploads/2024/04/protein-dot-blot-protocol-workflow-1024x576.webp)Protein dot blot workflow showing sample spotting on a membrane, blocking, primary and secondary antibody incubation, washing, and detection of circular spots.

- Prepare the protein samples at suitable concentrations. Different dilutions may be tested to obtain a clear signal. Include positive and negative controls.

- Cut the nitrocellulose or PVDF membrane according to the number of samples. Wet the nitrocellulose membrane with a suitable buffer. PVDF is first treated with methanol if required by the protocol.

- Apply a small volume of protein sample on the marked position of the membrane using a micropipette. A dot blot manifold can also be used. Avoid touching the membrane with the pipette tip.

- Allow the protein to bind with the membrane. The membrane may be dried after manual spotting or processed directly according to the protocol.

- Place the membrane in a suitable blocking solution. It blocks the remaining free protein-binding sites and prevents nonspecific antibody binding.

- Incubate the membrane with a primary antibody specific to the target protein. The antibody concentration must be optimized. Higher concentration may increase background, while very low concentration may produce a weak signal.

- Wash the membrane to remove unbound primary antibodies. Add a suitable labeled secondary antibody if indirect detection is used. Wash again to remove excess secondary antibody. This step is not required when a directly labeled primary antibody is used.

- Add the required detection reagent and visualize the dots using chemiluminescence, fluorescence or another suitable method. Antibody dilution, incubation time, blocking and washing conditions are adjusted according to the sample and detection system.

### DNA Dot Blot Protocol

- Extract the DNA from the sample and prepare the required concentration or serial dilutions. Include positive and negative controls. Standards may also be used for relative comparison.

- Denature the DNA if single-stranded DNA is required for probe hybridization. Denaturation separates the two complementary strands and exposes the target sequence for probe binding.

- Apply the prepared DNA on a suitable nucleic acid-binding membrane. Positively charged nylon membrane is commonly used. The samples can be spotted manually or using a dot blot manifold with controlled vacuum.

- Fix the DNA on the membrane using UV cross-linking or another suitable fixation method. The fixation method depends on the membrane used. Proper fixation is required to retain the DNA during washing.

- Prehybridize the membrane in a suitable hybridization solution. Add the labeled probe complementary to the target DNA sequence and allow it to hybridize. The probe and hybridization conditions are selected according to the target DNA.

- Wash the membrane under suitable stringency conditions to remove unbound and insufficiently matched probes. Washing temperature and salt concentration are adjusted according to the probe-target hybrid.

- Detect the bound probe using a suitable detection system and record the signals. The appearance of a positive dot indicates the presence of the complementary DNA sequence. Signal intensity can also be compared for relative target abundance using suitable standards, equal loading and controlled detection conditions.

![DNA and RNA dot blot workflows showing nucleic acid preparation, spotting and fixation on nylon membranes, labeled probe hybridization, washing, and detection.](https://biologynotesonline.com/wp-content/uploads/2024/04/dna-rna-dot-blot-hybridization-workflow-1024x576.webp)DNA and RNA dot blot workflows showing nucleic acid preparation, spotting and fixation on nylon membranes, labeled probe hybridization, washing, and detection.

### RNA Dot Blot Protocol

- Extract the RNA using RNase-free materials. RNA is easily degraded by ribonucleases. Use RNase-free tubes, water, pipette tips and other materials during sample preparation.

- Determine the quantity and quality of the extracted RNA. Prepare the required dilutions. The RNA may be denatured according to the protocol to reduce secondary structure and make the target sequence accessible.

- Prepare a suitable nucleic acid-binding membrane. Positively charged nylon membrane is commonly used for RNA dot blot. Mark the sample positions before loading.

- Apply the RNA sample directly on the membrane and allow it to bind. Fix the immobilized RNA using UV cross-linking or another suitable method according to the membrane and protocol.

- Prehybridize the membrane if required. Add a labeled complementary probe and incubate under suitable hybridization conditions. The probe binds with the specific target RNA sequence.

- Wash the membrane under suitable stringency conditions to remove unbound and insufficiently matched probes. Detect the bound probe using radioactive, fluorescent, enzyme-linked or another suitable detection system.

- Record the dot signals and compare them with positive, negative and loading controls. Differences in RNA loading must be considered during comparison. Total RNA staining, a suitable reference RNA or another validated method can be used for normalization.

In antibody-based RNA dot blot, the immobilized RNA is detected using antibodies specific to a particular RNA modification or structure. The bound antibody is then detected using a suitable antibody detection system. In hybridization-based RNA dot blot, a complementary labeled probe is used for the detection of a specific RNA sequence.

## Dot Blot Results and Interpretation

The dot blot result is observed from the signal produced at the position where each sample was spotted on the membrane. Depending on the detection system, the signal may be visible as colour, fluorescence or chemiluminescence. The membrane background and negative control are also checked during interpretation.

Positive: A distinct dot having signal above the negative control or established background is considered as a positive result. It indicates detection of the target in that sample.

Negative: No detectable dot, or signal remaining near the negative-control/background level. If the positive control also gives no signal, the negative sample result cannot be considered reliable.

The intensity of a dot may represent the relative amount of target. A sample containing more detectable target generally produces a stronger signal when equal amount of samples are loaded and the detection remains within its working range. Sample loading is important. Unequal loading of protein, deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) can itself produce differences in signal intensity.

For qualitative analysis, dots are mainly examined for presence or absence of the target signal. Dot blot can also be used for semiquantitative comparison. Here, signal intensities of different samples are compared after suitable loading normalization and correction of the background.

Actual quantitative analysis requires a validated assay. A calibration or standard curve prepared from known amounts of target is used and the unknown samples must fall within the linear range of detection. Replicates, proper controls, background subtraction and calibrated image analysis are also required. Different target molecules may not always produce the same signal response even when equal amounts are spotted.

![Illustrative dot blot membranes showing positive and negative controls, weak and diffuse dots, high background, saturation, and a schematic signal-response curve.](https://biologynotesonline.com/wp-content/uploads/2024/04/dot-blot-results-signal-interpretation-1024x576.webp)Illustrative dot blot membranes showing positive and negative controls, weak and diffuse dots, high background, saturation, and a schematic signal-response curve.

### Common Problems in Dot Blot Results

- Weak signal- It may occur due to a low amount of target, poor target recognition or unsuitable antibody/probe conditions. A signal very close to membrane background becomes difficult to interpret.

- Diffuse dots- The spotted sample may spread beyond its original area and form an irregular dot. Measurement of such signal becomes less reliable.

- High background- Insufficient blocking, excessive antibody, nonspecific binding, contaminants or inadequate washing can increase the membrane background. The difference between positive and negative dots then becomes less clear.

- Signal saturation- Excess sample or very high detection exposure can produce a saturated dot. After saturation, further increase of target does not produce a proportional increase in measured signal. Saturated dots cannot be used for quantitative comparison.

## Applications of Dot Blot

- Used to detect specific proteins and antigens in biological samples by antibody-based detection.

- Used for detection of specific antibodies in serum and for serological screening.

- Used to screen [hybridoma cultures](https://biologynotesonline.com/monoclonal-antibodies/) and identify antibody-producing clones.

- Used to detect specific deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) sequences by complementary probe hybridization.

- Used for comparison of relative amounts of proteins or nucleic acids present in different samples. Quantitative dot blot can also be used with suitable standards and validated assay conditions.

- Used for rapid screening of many samples in basic research and clinical testing.

## Advantages of Dot Blot

- It is a simple and rapid method because prior electrophoretic separation is not required.

- It can analyze many samples at the same time and is useful for high-throughput screening.

- Only a small amount of sample is required. It is useful when biological sample is limited or difficult to obtain.

- Dot blot is comparatively inexpensive and requires less equipment and reagents than conventional Western blot analysis.

- It can provide quick qualitative detection and semiquantitative comparison of target amounts. With proper calibration and validated conditions, quantitative dot blot analysis can also be performed.

- The technique can be used for both proteins and nucleic acids by changing the target-specific detection system.

## Limitations of Dot Blot

- It does not separate the sample molecules according to their molecular size. Protein isoforms, cleavage products and different modified forms may produce a combined dot signal.

- The specificity of dot blot depends on the antibody or nucleic acid probe used. Cross-reactivity can give nonspecific or false-positive signal.

- Precise quantification is difficult without proper calibration. High amount of target can saturate the signal. Very weak signals may also become difficult to distinguish from the membrane background.

- Small differences in sample loading and spotting can change the dot size and its intensity. This reduces reproducibility between samples or separate experiments.

- Insufficient blocking, poor washing or excessive antibody may produce a high membrane background and interfere with detection of weak target signals.

- Compared to an electrophoretic blot, dot blot provides less information about the target because only the final spot signal is obtained. For accurate protein identification, antibody specificity must be independently validated.

## Dot Blot Compared with Other Blotting Methods

![Comparison of dot blot and slot blot direct membrane spotting with Western blot gel electrophoresis and transfer, showing circular dots, rectangular slots, and protein bands.](https://biologynotesonline.com/wp-content/uploads/2024/04/dot-blot-vs-western-blot-vs-slot-blot-1024x576.webp)Comparison of dot blot and slot blot direct membrane spotting with Western blot gel electrophoresis and transfer, showing circular dots, rectangular slots, and protein bands.

### Dot Blot vs Western Blot

In dot blot, the sample is directly applied on the membrane and the particular target is detected from the resulting dot. No electrophoretic separation is done. In [Western blot](https://biologynotesonline.com/western-blot-protocol-principle-result/), proteins are first separated by gel electrophoresis according to their molecular size, transferred onto a membrane and then detected using specific antibodies.

Dot blot mainly gives information about the presence and relative amount of the target. Western blot gives a band at a particular position and also provides approximate molecular-mass information. This can help to distinguish the expected protein from proteins of different sizes, cleavage products or some other antibody-reactive proteins.

The dot blot procedure is shorter and many samples can be screened together. Western blot requires electrophoresis and transfer, so the procedure has more steps. It is more useful when protein size and band pattern are required for interpretation. Dot blot is commonly preferred for rapid screening or comparison of a large number of samples.

Neither dot blot nor Western blot is always more accurate. The result depends on antibody specificity, sample preparation, target abundance, detection range, controls and the actual purpose of the experiment. Western blot provides an additional size-based check, whereas dot blot avoids this separation completely.

### Dot Blot vs Slot Blot

Dot blot and slot blot follow almost the same basic principle. The sample is directly immobilized on a membrane without electrophoretic separation and the target is then detected by antibody binding or nucleic acid hybridization, depending on the assay. Their major difference is the geometry of sample application.

A dot blot forms a circular spot, whereas slot blot apparatus applies the sample as an elongated or rectangular area. The manifold used for loading is different. The rectangular geometry can change the distribution and area of the signal used for densitometric measurement, so uniform loading of each slot is required. It does not represent a different molecular detection mechanism.

FeatureDot BlotWestern BlotSlot BlotSample depositionSample is applied directly as a circular dot on the membrane.Protein sample is first loaded into a gel and the separated proteins are later transferred to membrane.Sample is applied directly through an elongated or rectangular slot.Electrophoretic separationNot required.Required before membrane transfer.Not required.Signal appearanceCircular dot or spot.One or more bands at particular positions on the membrane.Elongated or rectangular signal area.Size informationDoes not provide molecular-size information.Provides approximate molecular-mass information from the position of the protein band.Does not provide molecular-size information.Appropriate useRapid target detection, screening of many samples and relative comparison of target amounts.Protein identification where target detection together with molecular size and band pattern is required.Direct target detection and relative measurement where slot-shaped sample application is preferred.

## References

- Bergmans, H. E., & Gaastra, W. (1988). Dot-blot hybridization method. Methods in Molecular Biology, 4, 385–390. [https://doi.org/10.1385/0-89603-127-6:385](https://doi.org/10.1385/0-89603-127-6:385)

- Brown, T. (2001). Dot and slot blotting of DNA. Current Protocols in Molecular Biology, Chapter 2, Unit 2.9B. [https://doi.org/10.1002/0471142727.mb0209bs21](https://doi.org/10.1002/0471142727.mb0209bs21)

- Chen, T., & Lin, H.-K. (2024). Protocol to analyze the role of various metabolites in impacting global RNA 5-methylcytosine levels in cultured cells by dot blot. STAR Protocols, 5(1), 102815. [https://doi.org/10.1016/j.xpro.2023.102815](https://doi.org/10.1016/j.xpro.2023.102815)

- Chlebowski, A. C., & Kisby, G. E. (2020). Protocol for high-throughput screening of neural cell or brain tissue protein using a dot-blot technique with near-infrared imaging. STAR Protocols, 1(2), 100054. [https://doi.org/10.1016/j.xpro.2020.100054](https://doi.org/10.1016/j.xpro.2020.100054)

- Dowling, J. W., Smith, J. R., & Forero, A. (2024). Protocol for detection of in vitro R-loop formation using dot blots. STAR Protocols, 5(1), 102857. [https://doi.org/10.1016/j.xpro.2024.102857](https://doi.org/10.1016/j.xpro.2024.102857)

- Ghosh, R., Gilda, J. E., & Gomes, A. V. (2014). The necessity of and strategies for improving confidence in the accuracy of western blots. Expert Review of Proteomics, 11(5), 549–560. [https://doi.org/10.1586/14789450.2014.939635](https://doi.org/10.1586/14789450.2014.939635)

- Greenfield, E. A. (2021). Hybridoma screening by antibody capture: Dot blot. Cold Spring Harbor Protocols, 2021(11), pdb.prot103051. [https://doi.org/10.1101/pdb.prot103051](https://doi.org/10.1101/pdb.prot103051)

- Hnasko, T. S., & Hnasko, R. M. (2015). The Western blot. Methods in Molecular Biology, 1318, 87–96. [https://doi.org/10.1007/978-1-4939-2742-5_9](https://doi.org/10.1007/978-1-4939-2742-5_9)

- Hong, S., Lee, H.-A., Park, S.-H., & Kim, O. (2011). Sensitive and specific detection of Mycoplasma species by consensus polymerase chain reaction and dot blot hybridization. Laboratory Animal Research, 27(2), 141–145. [https://doi.org/10.5625/lar.2011.27.2.141](https://doi.org/10.5625/lar.2011.27.2.141)

- Litovchick, L. (2020). Blocking of immunoblots and incubation with antibodies: Procedure for immunoblots prepared with immunoprecipitated protein antigens. Cold Spring Harbor Protocols, 2020(6), pdb.prot098475. [https://doi.org/10.1101/pdb.prot098475](https://doi.org/10.1101/pdb.prot098475)

- Lu, J., Mai, F.-Y., Li, X.-Y., Yang, W.-T., Liang, J.-R., Li, X.-L., Guo, J., & Li, C.-G. (2026). Advances in protein dot blot: Principles, technical specifics, applications, and future perspectives. Frontiers in Molecular Biosciences, 13, 1768231. [https://doi.org/10.3389/fmolb.2026.1768231](https://doi.org/10.3389/fmolb.2026.1768231)

- Magi, B., & Liberatori, S. (2005). Immunoblotting techniques. Methods in Molecular Biology, 295, 227–254. [https://doi.org/10.1385/1-59259-873-0:227](https://doi.org/10.1385/1-59259-873-0:227)

- Mishra, V. (2022). Dot-blotting: A quick method for expression analysis of recombinant proteins. Current Protocols, 2(9), e546. [https://doi.org/10.1002/cpz1.546](https://doi.org/10.1002/cpz1.546)

- Norouzi, M., Zayeni, R., Singh, S., & Pardee, K. (2025). Cell-free dot blot as a practical and adaptable immunoassay platform for the detection of antibody response in human and animal sera. Journal of Visualized Experiments, 2025(219), e67973. [https://doi.org/10.3791/67973](https://doi.org/10.3791/67973)

- Old, J., Harteveld, C. L., Traeger-Synodinos, J., Petrou, M., Angastiniotis, M., & Galanello, R. (2012). Prevention of thalassaemias and other haemoglobin disorders: Volume 2: Laboratory protocols (2nd ed.). Thalassaemia International Federation. [https://www.ncbi.nlm.nih.gov/books/NBK190576/](https://www.ncbi.nlm.nih.gov/books/NBK190576/)

- Powers, J. M., Chang, X. L., Song, Z., & Nakai, H. (2018). A quantitative dot blot assay for AAV titration and its use for functional assessment of the adeno-associated virus assembly-activating proteins. Journal of Visualized Experiments, 2018(136), e56766. [https://doi.org/10.3791/56766](https://doi.org/10.3791/56766)

- Ramirez, P., Crouch, R. J., Cheung, V. G., & Grunseich, C. (2021). R-loop analysis by dot-blot. Journal of Visualized Experiments, 2021(167), e62069. [https://doi.org/10.3791/62069](https://doi.org/10.3791/62069)

- Sari, T. K., Gianopulos, K. A., & Nicola, A. V. (2020). Conformational change in herpes simplex virus entry glycoproteins detected by dot blot. Methods in Molecular Biology, 2060, 319–326. [https://doi.org/10.1007/978-1-4939-9814-2_18](https://doi.org/10.1007/978-1-4939-9814-2_18)

- Subramanian, S., Mahadevan, A., Satishchandra, P., & Shankar, S. K. (2016). Development of a dot blot assay with antibodies to recombinant “core” 14-3-3 protein: Evaluation of its usefulness in diagnosis of Creutzfeldt–Jakob disease. Annals of Indian Academy of Neurology, 19(2), 205–210. [https://doi.org/10.4103/0972-2327.176867](https://doi.org/10.4103/0972-2327.176867)

- Tian, G., Tang, F., Yang, C., Zhang, W., Bergquist, J., Wang, B., Mi, J., & Zhang, J. (2017). Quantitative dot blot analysis (QDB), a versatile high throughput immunoblot method. Oncotarget, 8(35), 58553–58562. [https://doi.org/10.18632/oncotarget.17236](https://doi.org/10.18632/oncotarget.17236)