Protein Block (ab64226)
Overview
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Product name
Protein Block -
Tested applications
Suitable for: IHC-P, ELISA, WBmore details -
General notes
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.
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.
6. Incubate with biotinylated secondary antibody (or 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 or 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
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Form
Liquid -
Storage instructions
Store at +4°C. -
Storage buffer
pH: 7.2
Preservative: 0.1% Sodium azide
Constituents: PBS, 0.5% Casein, 0.5% BSA -
Concentration information loading... -
Reagent notes
ab64226 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. -
Research areas
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Relevance
Protein Block is used with immunolabeling techniques for the reduction of non-specific background staining.
Associated products
Applications
The Abpromise guarantee
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.
| Application | Abreviews | Notes |
|---|---|---|
| IHC-P |
Use at an assay dependent dilution.
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| ELISA |
Use at an assay dependent dilution.
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| WB |
Use at an assay dependent dilution.
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| Notes |
|---|
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IHC-P
Use at an assay dependent dilution. |
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ELISA
Use at an assay dependent dilution. |
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WB
Use at an assay dependent dilution. |
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Protein Block (ab64226)
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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Protein Block (ab64226)Yu S.H., PLoS One 9 (12), Fig 7c. doi: 10.1371/journal.pone.0114649 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
References (8)
ab64226 has been referenced in 8 publications.
- Nishimori S et al. PTHrP targets HDAC4 and HDAC5 to repress chondrocyte hypertrophy. JCI Insight 4:N/A (2019). 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). PubMed: 29950590
- Ferreira-Gonzalez S et al. Paracrine cellular senescence exacerbates biliary injury and impairs regeneration. Nat Commun 9:1020 (2018). PubMed: 29523787
- Vasconcelos ACU et al. A symptomatic swelling of the upper lip. Oral Surg Oral Med Oral Pathol Oral Radiol 125:107-111 (2018). PubMed: 29248422
- Bostner J et al. Raptor localization predicts prognosis and tamoxifen response in estrogen receptor-positive breast cancer. Breast Cancer Res Treat 168:17-27 (2018). PubMed: 29128895
- Botti G et al. COX-2 expression positively correlates with PD-L1 expression in human melanoma cells. J Transl Med 15:46 (2017). PubMed: 28231855
- Skardal A et al. Bioprinted amniotic fluid-derived stem cells accelerate healing of large skin wounds. Stem Cells Transl Med 1:792-802 (2012). IHC-Fr . PubMed: 23197691
- Takeda K et al. Expression of LGR5, an intestinal stem cell marker, during each stage of colorectal tumorigenesis. Anticancer Res 31:263-70 (2011). PubMed: 21273608
