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.  

Ordering information for ELISA blocking buffers, kits and related products

Catalog # Name Size Price (USD) Qty
37545 SuperBlock™ (TBS) Blocking Buffer Dry Blend, 5 packs Each
182.65

Online exclusive

209.00
Save 26.35 (13%)
37581 SuperBlock™ (TBS) Blocking Buffer, 100 mL Each
52.65

Online exclusive

58.00
Save 5.35 (9%)
37535 SuperBlock™ (TBS) Blocking Buffer, 1 L Each
231.65

Online exclusive

260.00
Save 28.35 (11%)
37580 SuperBlock™ (PBS) Blocking Buffer, 100 mL Each
52.65

Online exclusive

58.00
Save 5.35 (9%)
37515 SuperBlock™ (PBS) Blocking Buffer, 1 L Each
223.65

Online exclusive

254.00
Save 30.35 (12%)
37518 SuperBlock™ (PBS) Blocking Buffer, 5 L Each
832.00
37516 SuperBlock™ T20 (PBS) Blocking Buffer, 1 L Each
250.65

Online exclusive

279.00
Save 28.35 (10%)
37536 SuperBlock™ T20 (TBS) Blocking Buffer, 1 L Each
251.65

Online exclusive

279.00
Save 27.35 (10%)
37584 Pierce™ Protein-Free (PBS) Blocking Buffer, 100 mL Each
51.65

Online exclusive

57.50
Save 5.85 (10%)
37572 Pierce™ Protein-Free (PBS) Blocking Buffer, 1 L Each
224.65

Online exclusive

253.00
Save 28.35 (11%)
37585 Pierce™ Protein-Free (TBS) Blocking Buffer, 100 mL Each
51.65

Online exclusive

57.50
Save 5.85 (10%)
37570 Pierce™ Protein-Free (TBS) Blocking Buffer, 1 L Each
228.65

Online exclusive

242.00
Save 13.35 (6%)
37573 Pierce™ Protein-Free T20 (PBS) Blocking Buffer, 1 L Each
244.65

Online exclusive

271.00
Save 26.35 (10%)
37571 Pierce™ Protein-Free T20 (TBS) Blocking Buffer, 1 L Each
249.65

Online exclusive

274.00
Save 24.35 (9%)
37578 StartingBlock™ (PBS) Blocking Buffer, 100 mL Each
52.65

Online exclusive

57.50
Save 4.85 (8%)
37538 StartingBlock™ (PBS) Blocking Buffer, 1 L Each
232.65

Online exclusive

262.00
Save 29.35 (11%)
37538X3 StartingBlock™ (PBS) Blocking Buffer 3 x 1 L
594.00
37579 StartingBlock™ (TBS) Blocking Buffer, 100 mL Each
51.65

Online exclusive

57.50
Save 5.85 (10%)
37542 StartingBlock™ (TBS) Blocking Buffer, 1 L Each
235.65

Online exclusive

263.00
Save 27.35 (10%)
37539 StartingBlock™ T20 (PBS) Blocking Buffer, 1 L Each
281.00
37543 StartingBlock™ T20 (TBS) Blocking Buffer, 1 L Each
250.65

Online exclusive

279.00
Save 28.35 (10%)
37582 Blocker™ Casein in PBS, 100 mL Each
49.65

Online exclusive

55.25
Save 5.60 (10%)
37528 Blocker™ Casein in PBS, 1 L Each
168.65

Online exclusive

191.00
Save 22.35 (12%)
37528X3 Blocker™ Casein in PBS 3 x 1 L
435.00
37583 Blocker™ Casein in TBS, 100 mL Each
49.65

Online exclusive

55.25
Save 5.60 (10%)
37532 Blocker™ Casein in TBS, 1 L Each
176.65

Online exclusive

191.00
Save 14.35 (8%)
37520 Blocker™ BSA (10X) in TBS, 125 mL Each
183.65

Online exclusive

206.00
Save 22.35 (11%)
37525 Blocker™ BSA (10X) in PBS, 200 mL Each
196.65

Online exclusive

222.00
Save 25.35 (11%)
37525X3 Blocker™ BSA (10X) in PBS 3 x 200 mL
502.00
37527 Fish Serum Blocking Buffer, 500 mL Each
229.65

Online exclusive

257.00
Save 27.35 (11%)
37527X3 Fish Serum Blocking Buffer, 3 x 500 mL Each
614.00
37530 Blocker™ BLOTTO in TBS Each
173.00
37565 Blocker™ FL Fluorescent Blocking Buffer (10X) Each
147.65

Online exclusive

161.00
Save 13.35 (8%)
37587 Pierce™ Clear Milk Blocking Buffer (10X) Each
187.65

Online exclusive

203.00
Save 15.35 (8%)
37575 Pierce™ Fast Blocking Buffer, 500 mL Each
282.65

Online exclusive

300.00
Save 17.35 (6%)
CNB0011 ELISA Buffer Kit Each
560.65

Online exclusive

594.00
Save 33.35 (6%)
900-K00 ABTS ELISA Buffer Kit, PeproTech® Each
256.00
900-T00 TMB ELISA Buffer Kit, PeproTech® Each
256.00
DS98200 ELISA Assay Buffer (5X) Each
208.65

Online exclusive

218.00
Save 9.35 (4%)
00-4202-56 eBioscience™ ELISA/ELISPOT Diluent (5X) Each
96.65

Online exclusive

104.00
Save 7.35 (7%)

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
106.65

Online exclusive

118.00
Save 11.35 (10%)
WB01 Wash Buffer (25X), 100 mL 100 ml
96.75
00-0400-46 eBioscience™ ELISA Wash Buffer, 1L packets Each
183.00
28382 BupH™ Carbonate-Bicarbonate Buffer Packs, 40 Packs Each
198.65

Online exclusive

221.00
Save 22.35 (10%)
28379 BupH™ Tris Buffered Saline Packs, 10 Packs Each
94.65

Online exclusive

107.00
Save 12.35 (12%)
28376 BupH™ Tris Buffered Saline Packs, 40 Packs Each
197.65

Online exclusive

220.00
Save 22.35 (10%)
28358 Pierce™ 20X TBS Buffer, 500 mL Each
110.65

Online exclusive

122.00
Save 11.35 (9%)
28374 BupH™ Modified Dulbecco's PBS Packs, 40 Packs Each
190.65

Online exclusive

222.00
Save 31.35 (14%)
28344 Pierce™ 20X Modified Dulbecco's PBS Buffer, 500 mL Each
122.65

Online exclusive

132.00
Save 9.35 (7%)
28346 Pierce™ 20X Modified Dulbecco's PBS Tween™ 20 Buffer, 500 mL Each
119.65

Online exclusive

133.00
Save 13.35 (10%)
28352 Pierce™ 20X PBS Tween™ 20 Buffer, 500 mL Each
110.65

Online exclusive

123.00
Save 12.35 (10%)
28360 Pierce™ 20X TBS Tween™ 20 Buffer, 500 mL Each
110.65

Online exclusive

123.00
Save 12.35 (10%)
SS03 ELISA Stop Solution, 25 mL 25 mL
71.65

Online exclusive

74.00
Save 2.35 (3%)
SS04 ELISA Stop Solution, 1 L 1 L
237.65

Online exclusive

248.00
Save 10.35 (4%)
N600 Stop Solution for TMB Substrates Each
99.65

Online exclusive

113.00
Save 13.35 (12%)

Ordering information for recommended cell lysis buffers

Catalog # Name Size Price (USD) Qty
FNN0011 Cell Extraction Buffer Each
177.65

Online exclusive

185.00
Save 7.35 (4%)
FNN0021 Cell Lysis Buffer II Each
179.65

Online exclusive

187.00
Save 7.35 (4%)
FNN0071 Tissue Extraction Reagent I Each
177.65

Online exclusive

187.00
Save 9.35 (5%)
FNN0081 Tissue Extraction Reagent II Each
185.00
78503 M-PER™ Mammalian Protein Extraction Reagent, 25 mL Each
89.65

Online exclusive

104.00
Save 14.35 (14%)
78501 M-PER™ Mammalian Protein Extraction Reagent, 250 mL Each
351.65

Online exclusive

390.00
Save 38.35 (10%)
78505 M-PER™ Mammalian Protein Extraction Reagent, 1 L Each
924.65

Online exclusive

1,016.00
Save 91.35 (9%)
78510 T-PER™ Tissue Protein Extraction Reagent Each
381.65

Online exclusive

426.00
Save 44.35 (10%)
87792 N-PER™ Neuronal Protein Extraction Reagent Each
282.65

Online exclusive

320.00
Save 37.35 (12%)

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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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      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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