Spark Red™ 718 anti-human HLA-A,B,C (Flexi-Fluor™) Antibody

Product Details

Verified Reactivity

Human, Cynomolgus, Rhesus

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 Red™ 718 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

Red Laser (633 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 sections^4,5, blocking^6,7, inhibition of NK cell-mediated lysis¹⁰, and activation^8,9. 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)

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

3105 View all products for this Gene ID

UniProt

View information about HLA-A on UniProt.org

Documentation

• Technical data sheet
• Certificate of Analysis
• Safety Data Sheet

Reviews

Related Protocols

• Cell Surface Flow Cytometry Staining Protocol

Related Pages & Pathways

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

Certificate of Analysis