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Blocking Solutions for WB and ELISA

Selection of Blocking reagents for WB and ELISA

Background

We have developed 4 types of Blocking Solutions for each field of Western blot and ELISA. Blocking is performed to minimize the non-specific binding of antigens or antibodies. The chemical property of each antigen, antibodies and the material where reaction occurs all of which are the cause of non-specific binding, blocking agent has to be selected to get maximum results. The blocking solutions for ELISA are desinged to long-term preservation of pre-coated plate.

Please try our blocking solutions if you use Skim milk for WB, or if you need storage of pre-coated ELISA plates.

Product Feature

*Ready-to-Use
*Optimized for WB or ELISA
*For WB, stronger signal can be obtained by using Skim milk 1)
*For ELISA, pre-coated plate can be stored for months 2)

1) Based on experiment in our laboratory
2) Except that antibody or antigen can not stored for long time

Selection of Blocking agents

Blocking Solutions have following main ingredients and characteristics ;

c-Block: 100% chemical components. Good for WB and antibody-detecting ELISA.
h-Block: Casein-based. Good for many applications.
k-Block: Degnatured casein-based. Shows stable results in many applications.
b-Block: BSA-based. Good for many applications.
Trial set: A trial kit composed of all the above 4 solutions. Good for screening of best blocking solution

A general rule is described. We strongly recommend to select the best one by screening using Tril Set.

For Western Bot, many reserachers seems to use Skim milk. Skim milk is not bad choice, but it very often gives you weak signal due to its strong blocking action. Below is the recommendated use of our bloking solutions.
1, To get higher signal(accept some background increase):k-Block, c-Block
2, To reduce background(accept some decrease of signal):b-Block, h-Block
3, To use as a standard blocking:h-Block, b-Block
4, To detect phospho-protein:c-Block, b-Block

For ELISA, below shows choice of blocking solutions depending on the ELISA system you use. All the solutions for ELISA are desinged for long term preservation, and dryed precoated plate can be stored for a few months after blocking at 4℃.
Antigen-detecting ELISA:
Direct method (detect antigen adsorbed on plate by detecting antibody):c-Block, b-Block, k-Block
Antibody-sandwich method (detect antigen by captured antibody by detect antibody):k-Block, b-Block, h-Block

Antibody-detecting ELISA:
Direct method (detect antibody captured by pre-coated antigen):c-Block, b-Block, k-Block
Antigen-sandwich method (detect antibody by captured antigen by detect antigen):k-Block, b-Block, h-Block Recommended to use together with Immunoshot Platinum

Example of experiment

Blocking Reagent -Chemically Defined- Protein free blocking reagent

Example 1


Example 2


A: skim milk, TBS-T
B: skim milk,IMMUNO SHOT(1&2)
C: skim milk,IMMUNO SHOT -Platinum-
D: Blocking Reagent-Chemically Defined、IMMUNO SHOT -Platinum-
(Data source: Kyoto University, Graduate School of Biostudies, Negisi laboratory, Hironori Kato)

See More Experimental examples

Product List

Product Name Cat# Quantity Price

Blocking Reagent for WB -Chemically Defined-

CSR-IS-CD-500W 500ML

¥ 8,000
$ 107
€ 80

h-Block-w

BEC-BCL-BKHW-01 500ML

¥ 8,000
$ 107
€ 80

k-Block-w

BEC-BCL-BKKW01 500ML

¥ 8,000
$ 107
€ 80

b-Block-w

BEC-BCL-BKBW-01 500ML

¥ 10,000
$ 134
€ 100

Blocking solution Trial set (Western)

BEC-BCL-BKSW-01 4*20ML

¥ 4,000
$ 54
€ 40

Blocking Reagent for ELISA -Chemically Defined-

CSR-IS-CD-500E 500ML

¥ 10,000
$ 134
€ 100

h-Block-e

BEC-BCL-BKHE-01 500ML

¥ 10,000
$ 134
€ 100

k-Block-e

BEC-BCL-BKKE-01 500ML

¥ 10,000
$ 134
€ 100

b-Block-e

BEC-BCL-BKBE-01 500ML

¥ 12,000
$ 160
€ 120

Blocking solution Trial set (ELISA)

BEC-BCL-BKSE-01 4*20ML

¥ 5,000
$ 67
€ 50

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.