Overview

  • Product name
    Protein Block
  • Tested applications
    Suitable for: IHC-P, ELISA, WBmore details
  • General notes

    Protein blocking buffer for serum-free blocking of non-specific antibody binding in IHC, ELISA and western blot. No mixing or dilution required.

    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.

    Find more IHC kits and reagents for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and mounting in the IHC kits and reagents guide.

     

    PROTOCOL:
    Immunohistochemistry
    Incubate tissue section for 5 minutes at room temperature prior to application of the primary antibody. After incubation rinse once in buffer.
    ELISA Add Protein Block to microtiter well and incubate for 2-10 minutes prior to addition of sample. Rinse and continue procedure.
    Chemiluminescent Blot Protein Block has been reported to be an effective blocker for this technique with incubation times of one hour at room temperature or overnight at 4°C.

Properties

  • Form
    Liquid
  • Storage instructions
    Store at +4°C.
  • Storage buffer
    pH: 7.40
    Preservative: 0.1% Sodium azide
    Constituents: 0.5% BSA, 0.5% Casein, PBS
  • 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
  • Relevance
    Protein Block is used with immunolabeling techniques for the reduction of non-specific background staining.

Applications

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.
ELISA Use at an assay dependent dilution.
WB Use at an assay dependent dilution.

Images

  • 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.

References

This product has been referenced in:
  • Botti G  et al. COX-2 expression positively correlates with PD-L1 expression in human melanoma cells. J Transl Med 15:46 (2017). Read more (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 . Read more (PubMed: 23197691) »
See all 3 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Question
Answer

ab64226 is the protein block used in ab80437. A link to ab64226 is below:

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Answer

Thank you for contacting us.
The lab sent me the following information:
1. What is the compositions of the protein block? Does it contain BSA?
ANSWER: PBS solution, pH 7.6, with 0.5% BSA, 0.5% casein and less than 0.1% sodium azide
2. Can the block be used in ICC?
ANSWER: The product has not been tested in immunocytochemistry applications.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thank you for contacting us.

We do not currently offer double staining IHC kits. Your staining conditions are challenging as, of the two common HRP chromogens, only DAB is alcohol insoluable. While of the common AP chromagens, only BCIP/NBT is insoluable. This means that to do double labeling you will have to do a sequential stain first using one detection system and then with the other.

I have collected a list, with links, of the reagents which you would need for this type of staining.

1) After de-parrafinizing and re-hydrating the samples, perform an antigen retrieval step treating with Cirate buffer pH6.0,https://www.abcam.com/10x-Citrate-Buffer-pH-6-0-ab64214.html, in the microwave or pressure cooker. After performing the antigen retrieval rinse in cold tap water for 10 minutes.

2) Rinse slides in TBST,https://www.abcam.com/20x-TBS-T-with-Tween-20-ab64250.html.

3) Performing a blocking step using protein block,https://www.abcam.com/Protein-Block-ab64226.html.

4) Incubate sections with your first primary antibody, for clarity I will assume the CD44 antibody, raised in mouse.

5) Wash with TBST.

6) Block endogenous peroxidases using aHydrogen Peroxide Blocking Reagent,https://www.abcam.com/Hydrogen-Peroxide-Blocking-Reagent-ab94666.html.

7) Incubate your tissue with a biotinylated secondary antibody, in this instance anti-Mouse IgG,https://www.abcam.com/Biotinylated-Goat-anti-Mouse-IgG-H-L-Ready-to-Use-ab64255.html.

8) Wash with TBST.

9) Detect your first antibody usingStreptavidin Peroxidase (Ready to Use)https://www.abcam.com/Streptavidin-Peroxidase-Ready-to-Use-ab64269.html. Note that this does not containa serum or BSA solution as diluent. Serum may contain biotin therefore competing Streptavidin binding with biotinylated secondary antibody, therefore reducing binding activity.

10) Apply your chromogen, DABsubtrate,https://www.abcam.com/DAB-Substrate-Kit-ab64238.html.

11) Wash with TBST.

12)Performing a second blocking step using protein block,https://www.abcam.com/Protein-Block-ab64226.html.

13)Wash with TBST.

14)Incubate sections with the your second primary antibody, this time ALDH1, raised in rabbit.

15)Wash with TBST.

16)Incubate your tissue with a biotinylated secondary antibody, in this instance anti-Rabbit IgG,https://www.abcam.com/Biotinylated-Goat-Anti-Rabbit-IgG-H-L-Ready-to-use-ab64256.html

17)Wash with TBST.

18)Detect your first antibody usingStreptavidin Alkaline Phosphatase (Ready to Use)https://www.abcam.com/Streptavidin-Alkaline-Phosphatase-Ready-to-Use-ab64268.html.

19) Apply your second chromogenAlkaline Phosphatase chromogen (BCIP/NBT) - Ready to Use,https://www.abcam.com/Alkaline-Phosphatase-chromogen-BCIP-NBT-Ready-to-Use-ab7468.html.

18) Wash with TBST, counterstain if desired.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Most of our antibodies will remain stable for 7 days at room temperature. For ab28364, anti-CD31, it should still be viable and work as expected. As for the protein block, we have no information on how long it will remain viable at room temperature, but I think it would still work for you, although I cannot guarantee that it will still work since it was not stored correctly. In future when you place orders, you can specify a delivery date so that you can avoid a delivery arriving when there will not be someone available to receive and store the products as required. If there is anything else I can do, please let me know.

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