Date published: 2026-4-5
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β-catenin (E-5): sc-7963. Immunoperoxidase staining of formalin fixed, paraffin-embedded human rectum tissue showing membr
β-catenin (E-5): sc-7963. Immunoperoxidase staining of formalin fixed, paraffin-embedded human rectum tissue showing membrane and cytoplasmic staining of glandular cells. Blocking reagent used: UltraCruz® Blocking Reagent: sc-516214.
β-catenin (E-5): sc-7963. Immunoperoxidase staining of formalin fixed, paraffin-embedded human rectum tissue showing membrane and cytoplasmic staining of glandular cells. Blocking reagent used: UltraCruz® Blocking Reagent: sc-516214.

UltraCruz® Blocking Reagent: sc-516214

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Datasheets
  • 100 ml ready-to-use solution for blocking and dilution
  • Plant protein derived universal antibody diluent and blocking reagent for use in immunohistochemistry (IHC), immunofluorescence (IF), Western blotting (WB), and for other research applications.
  • animal-free alternative to sera, BSA or non-fat dry milk
  • Designed to stabilize diluted antibody solutions and minimize background due to non-specific binding in different antibody-based research applications
  • The UltraCruz® Blocking Reagent is especially convenient in multiplex IHC (multiple antigen labeling) applications where antibodies from several different species are used
  • Highly recommended for use with ImmunoCruz® Antibody Conjugates and for Near-Infrared (NIR) Western blot detection methods, such as LI-COR/Odyssey® and other comparable systems
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Description
Datasheets and Protocols

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UltraCruz® Blocking Reagent is developed for use in various immunodetection techniques to minimize non-specific binding and enhance signal specificity. This reagent is a cornerstone in the protocols of immunohistochemistry (IHC), immunofluorescence (IF), and Western blotting (WB), where it serves as both an antibody diluent and a blocking agent. The absence of animal-derived proteins in its composition addresses potential issues of cross-reactivity and makes it an ideal alternative to traditional blocking solutions such as serum, bovine serum albumin (BSA), casein, or non-fat dry milk. The mechanism by which UltraCruz® Blocking Reagent operates involves saturating potential non-specific binding sites on the substrate or tissue, thereby preventing the primary and secondary antibodies used in these assays from binding to areas other than the target antigens. This results in a marked reduction in background noise, allowing for clearer and more precise visualization of the target proteins. Its efficacy in improving signal-to-noise ratio makes it particularly valuable in research that demands high fidelity in imaging and protein detection. Researchers utilize this reagent to ensure that the visualization of proteins is due to specific interactions rather than non-specific adsorption. Its utility spans across disciplines requiring precise antibody-antigen interactions, including molecular biology and biochemistry, where accurate detection and quantification of proteins are crucial. The formulation of UltraCruz® Blocking Reagent is tailored to enhance the clarity and reliability of experimental results, thus supporting advanced research applications in protein expression and molecular interaction studies.

UltraCruz® Blocking Reagent References:

  1. Generation of Insulin-Producing Cells from Canine Adipose Tissue-Derived Mesenchymal Stem Cells.  |  Teshima, T., et al. 2020. Stem Cells Int. 2020: 8841865. PMID: 33133196
  2. Simultaneous Isolation of Circulating Nucleic Acids and EV-Associated Protein Biomarkers From Unprocessed Plasma Using an AC Electrokinetics-Based Platform.  |  Hinestrosa, JP., et al. 2020. Front Bioeng Biotechnol. 8: 581157. PMID: 33224932
  3. TRPM3 Channels Play Roles in Heat Hypersensitivity and Spontaneous Pain after Nerve Injury.  |  Su, S., et al. 2021. J Neurosci. 41: 2457-2474. PMID: 33478988
  4. A myeloid-stromal niche and gp130 rescue in NOD2-driven Crohn′s disease.  |  Nayar, S., et al. 2021. Nature. 593: 275-281. PMID: 33789339
  5. Systems-Level Proteomics Evaluation of Microglia Response to Tumor-Supportive Anti-Inflammatory Cytokines.  |  Ahuja, S. and Lazar, IM. 2021. Front Immunol. 12: 646043. PMID: 34566949
  6. Chitosan and Hyaluronic Acid Nanoparticles as Vehicles of Epoetin Beta for Subconjunctival Ocular Delivery.  |  Silva, B., et al. 2022. Mar Drugs. 20: PMID: 35200680
  7. Deciphering Stem Cell From Apical Papilla-Macrophage Choreography Using a Novel 3-dimensional Organoid System.  |  Li, FC., et al. 2022. J Endod. 48: 1063-1072.e7. PMID: 35513088
  8. A High-Fat Diet Induces Low-Grade Cochlear Inflammation in CD-1 Mice.  |  Chan, J., et al. 2022. Int J Mol Sci. 23: PMID: 35563572
  9. BNIP3 Is Involved in Muscle Fiber Atrophy in Late-Onset Pompe Disease Patients.  |  Carrasco-Rozas, A., et al. 2022. Am J Pathol. 192: 1151-1166. PMID: 35605642
  10. In Vitro Model of Human Skeletal Muscle Tissue for the Study of Resident Macrophages and Stem Cells.  |  Hao, D., et al. 2022. Biology (Basel). 11: PMID: 35741457
  11. Does TLS Exist in Canine Mammary Gland Tumours? Preliminary Results in Simple Carcinomas.  |  Giambrone, G., et al. 2022. Vet Sci. 9: PMID: 36423077
  12. Protective effect of fatty acid amide hydrolase inhibitor URB597 and monoacylglycerol lipase inhibitor KML29 on renal ischemia-reperfusion injury via toll-like receptor 4/nuclear factor-kappa B pathway.  |  Çakır, M., et al. 2023. Int Immunopharmacol. 114: 109586. PMID: 36700769
  13. Topical ocular delivery of nanoparticles with epoetin beta in Wistar Hannover rats.  |  Silva, B., et al. 2023. Sci Rep. 13: 1559. PMID: 36707615

Ordering Information

Product NameCatalog #UNITPriceQtyFAVORITES

UltraCruz® Blocking Reagent

sc-516214
100 ml
$78.00

This product can be used as a blocking reagent in FCM?

Asked by: Lucia CJ
Thank you for your question. For flow cytometry we recommend using a high concentration of immunoglobulin that will bind to the Fc-receptors on cells. The blocking reagent should ideally be immunoglobulin from the species whose cells you are staining, such as normal mouse IgG: sc-2025.
Answered by: Tech Support Europe
Date published: 2023-06-15

SCBT, I've been using the Ultra-Cruz Blocking Reagent for IF staining, but I'm having issues with non-specific binding. I was hoping to check the age of the reagent, as it has no date on it (Lot: A1221). How long is the reagent good for after opening?

Asked by: cburk
Thank you for your question. We warranty this product for one year from date of purchase . Lot A1221 was manufactured December 2021. For assistance please contact technical service (800)-457-3801 or scbt@scbt.com.
Answered by: BlakeJ
Date published: 2023-04-21

How much volume of blocking buffer is required for 1 mini PVDF membrane and what time i should incubate?

Asked by: Sbayen
Thank you for your question . We recommend 5% and 1 hour at room temperature.
Answered by: BlakeJ
Date published: 2023-01-19

I have bought the UltraCruz® Blocking Reagent (sc-516214) but I cannot find any protocol how to use it. Do I need to dilute our antinbody in the blokker ( as we do with 1%BSA) or do we need to inclubate our tissue with the blokker separately?

Asked by: UMCG
Thank you for your question. It would be helpful if you could call us directly at 800-457-3801 ext .1, allowing for a more interactive discussion of this and other related questions
Answered by: Blake J
Date published: 2025-12-10

Can we use m-IgGκ BP-HRP: sc-516102 (Secondary antibody) with out the blocking buffer/reagent

Asked by: anjan1983
Thank you for your question. You can use the mouse IgG secondary reagent with any blocking buffer or reagent. I am not sure how you would use it without diluting it, but it could be possible.
Answered by: Tech Service
Date published: 2019-08-29

Dear sir  I am interested in your secondaries antibodies, but I knew from your website that your secondaries were not prepared in any live host, therefore there is high specificity and minimal background. Is there no need to use the blocking reagent

Asked by: Doaa
Thank you for your question. We still recommend using a blocking reagent for your research applications even when using our recombinant mouse IgG binding proteins in lieu of a conventional secondary antibody that is derived from a living host animal.
Answered by: Technical Support
Date published: 2017-12-19

Do I need to add detergent (exple: Triton X-100) to solubilize the membrane as with conventional Blocking sera to the UltraCruz® Blocking Reagent: sc-516214?

Asked by: Flo81
Thank you for your question. Yes, UltraCruz® Blocking Reagent: sc-516214 is a plant-based alternative to conventional blocking serums or buffers. It can be used in place of other blocking reagents. But if you typically use Triton X-100 in your protocols, then we would recommend to continue to do so as this buffer should not interfere with that procedure.
Answered by: Technical Support
Date published: 2018-07-12

Does UltraCruz® Blocking Reagent (sc-516214) contain glycine? What are the ingredients of this product? The datasheed does not provide any information. Thank you.

Asked by: Stark
Thank you for your question. Our UltraCruz® Blocking Reagent: sc-516214 is a proprietary buffer that is an animal-free alternative to sera, BSA or non-fat dry milk. The ingredients in the buffer are not provided as this is proprietary information. Please contact our Technical Service Department if you require additional information to purchase, or use this blocking buffer. Please call our European Support office toll-free at 00800 4573 8000, email europe@scbt.com, or contact us by live chat.
Answered by: Technical Support
Date published: 2019-06-04
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Rated 5 out of 5 by from Block reagentProtocol for immunohistochemistry caspase 8 and caspase 9
Date published: 2020-08-25
Rated 5 out of 5 by from great blocking buffer for IF on cellsvery nice working blocking buffer works better than BSA and animal serums
Date published: 2019-11-13
Rated 4 out of 5 by from good productsI bought the blocking reagent and is a good product making the research easier.
Date published: 2017-03-22
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UltraCruz ® Blocking Reagent is rated 4.7 out of 5 by 3.
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