Description
Key facts
Applications:WB, ELISA, IHC-PSee reactivity dataSee the reactivity data table below for information on validated species and application combinations.,
Form:LiquidSee storage information,
Storage buffer:pH: 7.2 - 7.6Preservative: 0.1% Sodium azideConstituents: PBS, 0.5% BSA, 0.5% Casein
Product details:
Protein Block has been developed to use with immunolabeling techniques for the reduction of nonspecific background staining while simultaneously reducing the handling of animal serums in the laboratory. The need to match species with the secondary antibody is eliminated due to the lack of normal serum in this product. Protein Block has been shown to be effective for immunohistochemical, ELISA, and blot and requires no mixing or diluting.
Protein blocking buffer ab64226 for serum-free blocking of non-specific antibody binding in IHC, ELISA and western blot. Ready to use. No mixing or dilution required.
For use in ELISA, add Protein Block to well and incubate for 2-10 minutes before addition of sample. Wash and continue procedure. For use in western blotting, incubate for one hour at room temperature or overnight at 4°C.
IHC protocol suitable for use with Protein Block ab64226:
For frozen sections, skip steps 1 and 2. For fluorescent IHC, skip step 3, incubate with fluorescent dye conjugated secondary at step 6, skip rest of steps, and mount with
anti-fade mounting medium
1. Deparaffinize and rehydrate formalin-fixed paraffin-embedded tissue section.
2. Use appropriate
antigen retrieval buffer or enzyme
(primary antibody dependent) to treat sections. Wash 3 times in buffer.
3. Add enough
hydrogen peroxide blocking solution
to cover the sections. Incubate for 10 minutes. Wash 2 times in buffer. If necessary, use
avidin biotin blocking
Apply Protein Block ab64226
(or normal serum from same species as secondary antibody)
and incubate for at least 30 minutes at room temperature
to block nonspecific background staining. Wash once in buffer.
5. Apply primary antibody in
antibody diluent
and incubate. Wash 4 times in buffer.
6. Incubate with
biotinylated secondary antibody
HRP polymer secondary antibody
and skip step 7). Wash 4 times in buffer.
7. Apply
streptavidin-HRP
and incubate for 10 minutes at room temperature.
8. Rinse 4 times in buffer. Place slide in
DAB substrate
AEC substrate
and incubate until desired color is achieved. Rinse 4 times in buffer.
9. Add enough drops of
hematoxylin
to cover the section. Incubate for 1 minute.
10. Rinse 7-8 times in tap water. Add
mounting medium
to cover the section.
Find complete IHC kits, and reagents for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and mounting in the
IHC kits and reagents guide
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C
