Product nameProtein Block
Tested applicationsSuitable for: IHC-P, ELISA, WBmore details
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.
4. Apply Protein Block ab64226 (or normal serum from same species as secondary antibody) and incubate for 5 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.
7. Apply streptavidin-HRP and incubate for 10 minutes at room temperature.
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.
Storage instructionsStore at +4°C.
Storage bufferpH: 7.40
Preservative: 0.1% Sodium azide
Constituents: PBS, 0.5% Casein, 0.5% BSA
Concentration information loading...
Reagent notesab64226 reduces 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. It has been shown to be effective for immunohistochemistry, ELISA, and blot and requires no mixing or diluting.
RelevanceProtein Block is used with immunolabeling techniques for the reduction of non-specific background staining.
Our Abpromise guarantee covers the use of ab64226 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use at an assay dependent dilution.|
|ELISA||Use at an assay dependent dilution.|
|WB||Use at an assay dependent dilution.|
ab64226 was used as pasr of the ab93677 anti-Mouse and Rabbit specific HRP (ABC) Detection IHC kit to perform immunohistochemical analysis of Human Tonsil tissue labeling Ki-67.
ab64226 was used in the mouse and rabbit specific HRP/DAB detection IHC Kit (ab64264) to visualize Immunohistochemical analysis staining CD163 in mouse soleus muscle section. Samples were incubated in rabbit CD163 antibody at 1:450 dilution.
This product has been referenced in:
- Nishimori S et al. PTHrP targets HDAC4 and HDAC5 to repress chondrocyte hypertrophy. JCI Insight 4:N/A (2019). Read more (PubMed: 30843886) »
- Salomão NG et al. BALB/c mice infected with DENV-2 strain 66985 by the intravenous route display injury in the central nervous system. Sci Rep 8:9754 (2018). Read more (PubMed: 29950590) »