Overview of ELISA buffers and reagents

ELISA buffers and reagents are important components to develop excellent assay performance for high sensitivity, low background, and blocking non-specific binding. Select from a wide variety of easy-to-use and reliable reagents including blocking buffers, coating buffers, wash buffers, and recommended cell lysis buffers for conducting successful ELISA assays. Or answer a few questions in our ELISA Builder and receive a full list of components required for designing your ELISA experiment.

Find ELISA buffers and reagents

ELISA buffers and reagents selection guide

Complete ELISA buffer kits are available based on substrate type. See all ELISA kits and buffers below.

Product nameComponents, benefitsSubstrate typeQuantityCat. No
 ELISA Buffer KitIncludes coating buffer A and B, assay buffer, wash buffer, TMB and stop solution TMB, HRP1 Kit*CNB0011
ABTS ELISA Buffer KitIncludes PBS, wash buffer, blocking buffer, diluent, ABTS substrate, ELISA microplates, sealing filmABTS, HRP1 Kit*900-K00
TMB ELISA Buffer KitIncludes PBS, wash buffer, blocking buffer, diluent, TMB substrate, ELISA microplates, sealing film, stop solution TMB, HRP1 Kit*900-T00
ELISA Assay Buffer (5X) Blocking reagent for ELISA plates, diluent for ELISA standards, samples, detection antibody, and HRP conjugate. 200 mLDS98200
ELISA/ELISPOT Diluent (5X)100 mL00-4202-56

*One kit provides sufficient material for 10 ELISA plates.

Learn more about these products in the section below under ELISA coating and wash buffers.

* Supplied as 30X concentrated solution
* Supplied as 25X concentrated solution
***Contains 0.05 % Tween-20, each pack yields 1L of 1X Wash Buffer

Complete ELISA buffer kits are available based on substrate type. See all ELISA kits and buffers below.

Product nameComponents, benefitsSubstrate typeQuantityCat. No
 ELISA Buffer KitIncludes coating buffer A and B, assay buffer, wash buffer, TMB and stop solution TMB, HRP1 Kit*CNB0011
ABTS ELISA Buffer KitIncludes PBS, wash buffer, blocking buffer, diluent, ABTS substrate, ELISA microplates, sealing filmABTS, HRP1 Kit*900-K00
TMB ELISA Buffer KitIncludes PBS, wash buffer, blocking buffer, diluent, TMB substrate, ELISA microplates, sealing film, stop solution TMB, HRP1 Kit*900-T00
ELISA Assay Buffer (5X) Blocking reagent for ELISA plates, diluent for ELISA standards, samples, detection antibody, and HRP conjugate. 200 mLDS98200
ELISA/ELISPOT Diluent (5X)100 mL00-4202-56

*One kit provides sufficient material for 10 ELISA plates.

Learn more about these products in the section below under ELISA coating and wash buffers.

* Supplied as 30X concentrated solution
* Supplied as 25X concentrated solution
***Contains 0.05 % Tween-20, each pack yields 1L of 1X Wash Buffer

ELISA blocking buffers

SuperBlock Blocking Bufferis a protein-based formulation that provides fast and effective blocking for ELISA, immunohistochemistry (IHC) and western blot analysis. It does not contain any immunoglobulins, albumin, or endogenous biotin, is compatible with streptavidin system, and yields a high signal-to-noise ratio. It may be used as a protein stabilizer for drying antigen or antibody-coated microplates.

SuperBlock Blocking Buffer is available in powder form and a ready-to-use solution. See selection guide (Tab 1) to choose for optimal reagent for your experiment.  

Protein-Free Blocking Buffer contains a protein-free compound and is highly effective at blocking ELISA microplates, membranes, and glass slide arrays. These buffers are free of biotin and are compatible with antibodies and avidin/biotin systems. They can be used to minimize or eliminate cross-reactivity associated with protein-blocking buffers and interference from glycosylation.

Protein-Free Blocking Buffers are optimized and validated in many protein methods to provide high signal-to-noise ratio and are supplied in a convenient ready-to-use format. See Figure 1 for comparison of Protein-Free Blocking Buffers and selection guide (Tab 1) to choose the suitable buffer for your experiment.

StartingBlock Blocking Buffer contains a single purified protein for instant blocking of western blot and ELISA assays especially chemiluminescent and fluorescent-based. These buffers provide broad compatibility with a wide range of antibodies and antibody combinations and are often used as a replacement of milk-based blocking buffers exhibiting high background or masking antigen-antibody binding. The blocker is serum-, albumin- and biotin-free and therefore is compatible with biotin/avidin systems.

StartingBlock Blocking Buffers are available as ready-to-use solution in PBS or TBS with and without Tween-20. See selection guide (Tab 1) to choose the best reagent for your experiment.  

Figure 1. Performance comparison between our Protein-Free Blocking Buffer and other commercial alternatives. Multiplex protein arrays were created by spotting 12 cytokine capture antibodies per well of 96-well microplates. The ELISA plates were then blocked with the indicated blocking buffers, probed with HRP-conjugated secondary antibody (no samples) and the background for each well was determined by chemiluminescence. Error bars represent the standard deviation for triplicate microplate wells. The graph shows that our Protein-Free Blocking Buffer exhibits less background than other commercial alternatives.

Blocker Caseincontains a single purified protein for blocking excess binding sites in ELISA, western blot, IHC, and nucleic acid detection methods. It provides fewer chances of cross-reactions with assay components and is a good alternative when high background or antigen-antibody masking is seen with non-fat milk blockers.

Blocker Casein is formulated in either PBS or TBS with 1% (w/v) casein and is available in a ready-to-use format. See selection guide (Tab 1) to choose this reagent for your experiment.  

Blocker BSA contains a single purified protein for blocking excess binding sites in western blot and ELISA, especially for targeting phospho-proteins. It provides fewer chances of cross-reactions with assay components and is a good alternative to non-fat milk blockers. It is devoid of endogenous biotin and is compatible with biotin/avidin systems. It may be used as a protein stabilizer for drying antigen or antibody-coated microplates, or used a diluent for storing used antibody solutions.

Blocker BSA is available in a ready-to-use format in either PBS or TBS with 10% (w/v) bovine serum albumin (BSA). See selection guide (Tab 1) to choose this reagent for your experiment.  

Fish Serum Blocking Buffer contains steelhead salmon serum and is an ideal blocking agent in ELISA, western blotting, immunohistochemistry, and other immunochemical detection methods involving mammalian samples, resulting in little to no background.  In addition, it is effective at stabilizing solutions and samples used for antibody-binding interactions and may be used as a diluent for antibodies.

Fish Serum Blocking Buffer is filtered and stabilized in PBS for compatibility with most assay systems and is supplied in a ready-to-use format. See selection guide (Tab 1) to choose this reagent for your experiment.  

ELISA coating and wash buffers

ELISA Phosphate Coating Buffer (Cat. No. CB07100) and ELISA Carbonate Coating Buffer (Cat. No. CB01100) are intended for use as a plate coating buffer for Invitrogen Antibody Pairs or for developing a sandwich ELISA.

ELISA Wash Buffer (Cat. No. N503) is available in either dried powder or concentrated liquid formulation and contains 0.05% Tween-20. See all ELISA wash buffers under selection guide (Tab 3).

Carbonate-Bicarbonate Buffer (pH 9.4) (Cat. No. 28382) is used for coating or binding capture-antibodies and other proteins onto ELISA plates and other plate assays. It is available in dry-blended powder form.

Tris-Buffered Saline (pH 7.2) (Cat. No. 28379) is used to prepare Tris-NaCl buffered saline (TBS) for antibody diluent and wash buffer in ELISA, western blotting, and general immunoassay procedures. It is available in either dry-blended powder form or concentrated stock solution. See all pack sizes under selection guide (Tab 3). 

Modified Dulbecco's PBS (Cat. No. 28374) and 20X Modified Dulbecco's PBS Tween-20 (Cat. No. 28346) are used to prepare physiological sodium- and potassium-phosphate buffered saline (D-PBS or D-PBS-T) for ELISA and western blot diluents and wash buffers. They are available in either dry-blended powder form or concentrated stock solution. See all pack sizes under selection guide (Tab 3). 

PBS Tween-20 (Cat. No. 28352) andTBS Tween-20 (Cat. No. 28360) are used to prepare PBS-Tween (PBS-T) or TBS-Tween (TBS-T) wash buffers or blocking buffer diluent for ELISA, western blotting, and other immunoassays. They are available as 20X concentrated stock solution.

Videos and webinars

How to use Invitrogen ELISA kits for protein analysis

View the instructional video demonstrating the workflow of a typical ELISA experiment using Invitrogen ELISA kits and reagents. Please make sure to carefully read and review your ELISA kit’s specific instruction sheet, located inside the package, for detailed information before beginning the procedure.

Additional ELISA products

Resources

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Immunoassay Research Areas

Explore our immunoassays for different research areas, including immune-oncology, neurobiology, isotyping and immunoglobulin characterization, hallmarks of cancer, car t cells, and more.

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For Research Use Only. Not for use in diagnostic procedures.

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