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Spark Blue™ 574 anti-human HLA-A,B,C (Flexi-Fluor™) Antibody

Pricing & Availability
Clone
W6/32 (See other available formats)
Regulatory Status
RUO
Other Names
Major Histocompatibility Class I, MHC class I
Isotype
Mouse IgG2a, κ
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Product Citations
publications
Compare all formats See Spark Blue™ 574 spectral data
Cat # Size Price Quantity Save
285187 50 µg 316€
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Description

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.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Reported Reactivity
African Green, Baboon, Cat, Cow, Chimpanzee
Antibody Type
Monoclonal
Host Species
Mouse
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography and conjugated with Spark Blue™ 574 under optimal conditions.
Concentration
0.2 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC
Recommended Usage

Flexi-Fluors™ are provided at a standard 0.2 mg/mL concentration. We recommend titrating this reagent to determine the optimal concentration for each application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL. We recommend testing a range of concentrations starting from 10 µg/mL.

For example, make five 1:1 serial dilutions of the 0.2 mg/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/mL) to test six concentrations -- 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.

Excitation Laser
Blue Laser (488 nm)
Application Notes

Clone W6/32 recognizes residues in the N terminus of the human ß2-microglobulin molecule21.

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 filtered) is recommended for functional assays. For highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 311428) with a lower endotoxin limit than standard LEAF™ purified antibodies (Endotoxin < 0.01 EU/µg).

Additional Product Notes

For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.

Application References

(PubMed link indicates BioLegend citation)
  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.
  18. Coppieters KT, et al. 2012. J. Exp. Med. 209:51. (epitope)
  19. Crivello P, et al. 2013. Hum Immunol. 22:100. PubMed
  20. Jung Y, et al. 2015. Mol Cancer Res. 13:197. PubMed
  21. Shields MJ. Ribaudo RK. 1998. Tissue Antigens. 51(5):567-70. (epitope)
RRID
AB_3662527 (BioLegend Cat. No. 285187)

Antigen Details

Structure
Ig superfamily
Distribution

All nucleated cells

Function
Antigen presentation
Ligand/Receptor
CD3/TCR, CD8
Biology Area
Immunology, Innate Immunity
Molecular Family
MHC Antigens
Antigen References

1. Barclay AN, et al. Eds. 1993. The Leukocyte Antigen FactsBook. Academic Press Inc. San Diego.

Gene ID
NA
UniProt
View information about HLA-A on UniProt.org

Related FAQs

What are Flexi-Fluors?

Flexi-Fluors are rapidly made-to-order conjugated antibodies. The technology, manufacturing processes, and specifications used to create Flexi-Fluors are the same as our regular catalog products. However, the optimal concentration and performance of each Flexi-Fluor must be determined by the customer. 

How quickly will I receive my order?

We aim to ship Flexi-Fluors within 2-3 weeks of receipt of your order. However, depending on your location, shipping times may vary. 

How are Flexi-Fluors different from regular catalog products?

Flexi-Fluors are made on demand, specifically for you. Flexi-Fluors are manufactured using the same high-quality standards, and specifications as other catalog products. For faster delivery, Flexi-Fluors are not tested by flow cytometry to determine optimal concentrations or evaluate performance. This testing needs to be performed by the customer.

How do I determine the optimal concentration for using my Flexi-Fluor? How should I titrate my antibody?

Flexi-Fluors are provided at a standard 0.2 mg/mL concentration. We recommend that you titrate your antibody to determine the optimal concentration to use for your application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL volume. We recommend that you test a range of concentrations starting from 10 µg/mL. 

 

For example, make five 1:1 serial dilutions of your 0.2 mg/mL antibody.  Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/ml) to test six concentrations - 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration. 

I can’t find the antibody-dye combination that I need. When will it be available?

We continuously update our catalog, introducing scores of new products every month. Please get in touch with our Technical Service team for an update on new products or recommendations for suitable alternatives to complete your panel. Or contact Custom Solutions to inquire about our affordable custom conjugation services. 

I need help to validate the performance of my Flexi-Fluor. Who should I contact?

Please get in touch with Technical Service for assistance. 

Can I order more than 50 μg of a Flexi-Fluor?

Yes, you can order multiple vials of the same Flexi-Fluor products.  We cannot guarantee, however, that these vials will be bottled from the same lot. For bulk single-lot orders, contact our Custom Solutions team.

What is the expiration date of my Flexi-Fluor?

Expiration dates can be found on the vial label or by using our CoA lookup tool. 

Other Formats

View All HLA-A,B,C Reagents Request Custom Conjugation
Description Clone Applications
APC anti-human HLA-A,B,C W6/32 FC
FITC anti-human HLA-A,B,C W6/32 FC
PE anti-human HLA-A,B,C W6/32 FC
PE/Cyanine5 anti-human HLA-A,B,C W6/32 FC
Purified anti-human HLA-A,B,C W6/32 FC,Activ,Block,IHC-F,IP,WB
Alexa Fluor® 488 anti-human HLA-A,B,C W6/32 FC
Alexa Fluor® 647 anti-human HLA-A,B,C W6/32 FC
Pacific Blue™ anti-human HLA-A,B,C W6/32 FC
PerCP anti-human HLA-A,B,C W6/32 FC
APC/Cyanine7 anti-human HLA-A,B,C W6/32 FC
PerCP/Cyanine5.5 anti-human HLA-A,B,C W6/32 FC
Ultra-LEAF™ Purified anti-human HLA-A,B,C W6/32 FC,Activ,Block,IHC-F,IP,WB
PE/Cyanine7 anti-human HLA-A,B,C W6/32 FC
Brilliant Violet 510™ anti-human HLA-A,B,C W6/32 FC
Alexa Fluor® 700 anti-human HLA-A,B,C W6/32 FC
PE/Dazzle™ 594 anti-human HLA-A,B,C W6/32 FC
Biotin anti-human HLA-A,B,C W6/32 FC
Brilliant Violet 605™ anti-human HLA-A,B,C W6/32 FC
APC/Fire™ 750 anti-human HLA-A,B,C W6/32 FC
TotalSeq™-A0058 anti-human HLA-A,B,C W6/32 PG
TotalSeq™-C0058 anti-human HLA-A,B,C W6/32 PG
TotalSeq™-B0058 anti-human HLA-A,B,C W6/32 PG
Spark NIR™ 685 anti-human HLA-A,B,C W6/32 FC
TotalSeq™-D0058 anti-human HLA-A,B,C W6/32 PG
GMP Ultra-LEAF™ Purified anti-human HLA-A,B,C SF W6/32 FC
GMP Ultra-LEAF™ Biotin anti-human HLA-A,B,C SF W6/32 FC
Spark UV™ 387 anti-human HLA-A,B,C W6/32 FC
Spark Red™ 718 anti-human HLA-A,B,C (Flexi-Fluor™) W6/32 FC
Spark Blue™ 574 anti-human HLA-A,B,C (Flexi-Fluor™) W6/32 FC
Spark Blue™ 550 anti-human HLA-A,B,C (Flexi-Fluor™) W6/32 FC
Go To Top Version: 1    Revision Date: 09.06.2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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.

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