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The antibody was purified by affinity chromatography, and conjugated with Pacific Blue™ under optimal conditions. The solution is free of unconjugated Pacific Blue™.
Concentration:
0.5 mg/ml
Storage & Handling:
The antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is ≤ 1..0 μg per 106 cells in 100 μl volume or 100 μl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.
* Pacific Blue™ has a maximum emission of 455 nm when it is excited at 405 nm. Prior to using Pacific Blue™ conjugate for flow cytometric analysis, please verify your flow cytometer's capability of exciting and detecting the fluorochrome. ** Pacific Blue™ is a registered trademark of Molecular Probes, Inc. Pacific Blue™ dye antibody conjugates are sold under license from Molecular Probes, Inc. for research use only, except for use in combination with microarrays and high content screening, and are covered by pending and issued patents.
The W6/32 monoclonal antibody reacts with a non-polymorphic epitope of human major histocompatibility complex (MHC) class I antigens, HLA-A, B, C. Additional reported applications (for the relevant formats) include: immunoprecipitaton2, Western blotting (non-reducing)3, immunohistochemical staining of acetone-fixed frozen tissue sections4,5, blocking6,7, inhibition of NK cell-mediated lysis10, and activation8,9. Clone W6/32 has been reported not to be suitable for immunohistochemistry on paraffin sections17. The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for functional assays (Cat. No. 311412).
Application References:
1. Darrow TL, et al. 1989. J. Immunol. 142:3329. 2. Stern P, et al. 1987. J. Immunol. 138:1088. 3. Tran TM, et al. 2001. Immunogenetics 53:440. 4. Barbatis C, et al. 1981. Gut 22:985. 5. Ayyoub M, et al. 2004. Cancer Immunity 4:7. 6. DeFelice M, et al. 1990. Cell. Immunol. 126:420. 7. Fayen J, et al. 1998. Int. Immunol. 10:1347. 8. Turco MC, et al. 1988. J. Immunol. 141:2275. 9. Geppert TD, et al. 1989. J. Immunol. 142:3763. 10. Wooden SL, et al. 2005. J. Immunol. 175:1383. 11. Nagano M, et al. 2007. Blood 110:151. 12. McLoughlin RM,et al.2008. J. Immunol. 181:1323. PubMed 13. Takahara M, et al.2008. J. Leukoc. Biol. 83:742. PubMed 14. Lunemann A, et al.2008. J. Immunol. 181:6170. PubMed 15. Laing BJ, et al. 2010. J. Thorac Cardiovasc Surg. 139:1402. PubMed 16. Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC) 17. Vambutas A, et al. 2000. Clin. Diagn. Lab. Immun. 7:79.
Human peripheral blood lymphocytes stained with W6/32 Pacific Blue™
MHC class I antigens associated with β2-microglobulin are expressed by all human nucleated cells. MHC class I molecules are involved in presentation of antigens to CD8+ T cells. They play an important role in cell-mediated immune responses and tumor surveillance.
Other Names:
Major Histocompatibility Class I, MHC class I
Structure:
Ig superfamily
Distribution:
All nucleated cells
Function:
Antigen presentation
Ligand Receptor:
CD3/TCR, CD8
Antigen References:
1. Barclay AN, et al. Eds. 1993. The Leukocyte Antigen FactsBook. Academic Press Inc. San Diego.
*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/terms). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.
This data display is provided for general comparisons between formats. Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors. If you need assistance with selecting the best format contact our expert technical support team.
APC anti-human HLA-A,B,C
Human peripheral blood lymphocytes stained with W6/32 APC
FITC anti-human HLA-A,B,C
Human peripheral blood lymphocytes stained with W6/32 FITC
PE anti-human HLA-A,B,C
Human peripheral blood lymphocytes stained with W6/32 PE
PE/Cy5 anti-human HLA-A,B,C
Human peripheral blood lymphocytes stained with W6/32 PE/Cy5
Purified anti-human HLA-A,B,C
HEK293 cells were transfected with RelA or empty vector and 24hrs later cell extracts harvested using a 1% CHAPS lysis buffer. Extracts were resolved by non-denaturing, non-reducing electrophoresis, transferred to nitrocellulose, and probed with a 1:500 dilution purified W6/32 . Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system. These data document that MHC class I was upregulated in cells constitutively expressing RelA. (Data was provided by Dr. Ezra Burstein, University of Michigan Medical School, Ann Arbor, MI).
Human peripheral blood lymphocytes stained with purified W6/32, followed by anti-mouse IgG FITC
LEAF™ Purified anti-human HLA-A,B,C
Human peripheral blood lymphocytes stained with LEAF™ purified W6/32, followed by anti-mouse IgG FITC
Alexa Fluor® 488 anti-human HLA-A,B,C
Human peripheral blood lymphocytes stained with W6/32 Alexa Fluor® 488
Alexa Fluor® 647 anti-human HLA-A,B,C
Human peripheral blood lymphocytes stained with W6/32 Alexa Fluor® 647
Pacific Blue™ anti-human HLA-A,B,C
Human peripheral blood lymphocytes stained with W6/32 Pacific Blue™
PerCP anti-human HLA-A,B,C
Human peripheral blood lymphocytes, monocytes and granulocytes stained with PerCP-conjugated W6/32 (bottom) and mIgG2a, k isotype control (top)
APC/Cy7 anti-human HLA-A,B,C
Human peripheral blood lymphocytes were stained with HLA-A,B,C (clone W6/32) APC/Cy7 (filled histogram) or mouse IgG2a APC/Cy7 isotype control (open histogram).
PerCP/Cy5.5 anti-human HLA-A,B,C
Human peripheral blood lymphocytes were stained with HLA-A,B,C (clone W6/32) PerCP/Cy5.5 (filled histogram) or mouse IgG2a PerCP/Cy5.5 isotype control (open histogram).
