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.

Ordering information for ELISA coating, wash buffers and stop solution

Catalog # Name Size Price (USD) Qty
CB07100 ELISA Phosphate Coating Buffer Each -
CB01100 ELISA Carbonate Coating Buffer Each -
N503 ELISA Wash Buffer (30X) Each
112.65

Online Exclusive

124.00
Save 11.35 (9%)
WB01 Wash Buffer (25X), 100 mL 100 ml
102.00
00-0400-46 eBioscience™ ELISA Wash Buffer, 1L packets Each
192.00
28382 BupH™ Carbonate-Bicarbonate Buffer Packs, 40 Packs Each
207.65

Online Exclusive

231.00
Save 23.35 (10%)
28379 BupH™ Tris Buffered Saline Packs, 10 Packs Each
99.65

Online Exclusive

112.00
Save 12.35 (11%)
28376 BupH™ Tris Buffered Saline Packs, 40 Packs Each
206.65

Online Exclusive

230.00
Save 23.35 (10%)
28358 Pierce™ 20X TBS Buffer, 500 mL Each
115.65

Online Exclusive

127.00
Save 11.35 (9%)
28374 BupH™ Modified Dulbecco's PBS Packs, 40 Packs Each
199.65

Online Exclusive

232.00
Save 32.35 (14%)
28344 Pierce™ 20X Modified Dulbecco's PBS Buffer, 500 mL Each
128.65

Online Exclusive

138.00
Save 9.35 (7%)
28346 Pierce™ 20X Modified Dulbecco's PBS Tween™ 20 Buffer, 500 mL Each
125.65

Online Exclusive

139.00
Save 13.35 (10%)
28352 Pierce™ 20X PBS Tween™ 20 Buffer, 500 mL Each
116.65

Online Exclusive

129.00
Save 12.35 (10%)
28360 Pierce™ 20X TBS Tween™ 20 Buffer, 500 mL Each
116.65

Online Exclusive

129.00
Save 12.35 (10%)
SS03 ELISA Stop Solution, 25 mL 25 mL
75.65

Online Exclusive

77.75
Save 2.10 (3%)
SS04 ELISA Stop Solution, 1 L 1 L
249.65

Online Exclusive

260.00
Save 10.35 (4%)
N600 Stop Solution for TMB Substrates Each
104.65

Online Exclusive

119.00
Save 14.35 (12%)

Ordering information for recommended cell lysis buffers

Catalog # Name Size Price (USD) Qty
FNN0011 Cell Extraction Buffer Each
185.65

Online Exclusive

193.00
Save 7.35 (4%)
FNN0021 Cell Lysis Buffer II Each
187.65

Online Exclusive

195.00
Save 7.35 (4%)
FNN0071 Tissue Extraction Reagent I Each
185.65

Online Exclusive

195.00
Save 9.35 (5%)
FNN0081 Tissue Extraction Reagent II Each
193.00
78503 M-PER™ Mammalian Protein Extraction Reagent, 25 mL Each
93.65

Online Exclusive

109.00
Save 15.35 (14%)
78501 M-PER™ Mammalian Protein Extraction Reagent, 250 mL Each
368.65

Online Exclusive

408.00
Save 39.35 (10%)
78505 M-PER™ Mammalian Protein Extraction Reagent, 1 L Each
966.65

Online Exclusive

1,062.00
Save 95.35 (9%)
78510 T-PER™ Tissue Protein Extraction Reagent Each
398.65

Online Exclusive

445.00
Save 46.35 (10%)
87792 N-PER™ Neuronal Protein Extraction Reagent Each
294.65

Online Exclusive

334.00
Save 39.35 (12%)

Videos and webinars

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    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.

    Tips and tricks for successful ELISA assay

    Explore our webinar around ELISA best practices for protein quantification. Learn how to analyze and interpret ELISA data, identify how to troubleshoot your ELISA experiment with useful tips and tricks to help you get reliable and accurate results.

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    For a complete set of ELISA reagents, Invitrogen ELISA Buffer Kit (Cat. No. CNB0011) includes 2 Coating Buffers (pH 7.4 and pH 9.4), 5X Assay Buffer (blocking reagent and diluent), 25X Wash Buffer, Stabilized TMB, and Stop Solution. Pair this kit with Matched Antibody Pair kits that contain matched antibody pairs, standards and Streptavidin-HRP conjugate.

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