Biolegend, 285168, Spark Blue™ 574 anti-human CD62L (Flexi-Fluor™) Antibody, 50μg

CD62L is a 74-95 kD single chain type I glycoprotein referred to as L-selectin or LECAM-1. It is expressed on most peripheral blood B cells, subsets of T and NK cells, monocytes, granulocytes, and certain hematopoietic malignant cells. CD62L binds to carbohydrates present on certain glycoforms of CD34, glycam-1, and MAdCAM-1 and with a low affinity to anionic oligosaccharide sequences related to sialylated Lewis X (sLex, CD15s) through its C-type lectin domain. CD62L is important for the homing of naïve lymphocytes to high endothelial venules in peripheral lymph nodes and Peyer's patches. It also plays a role in leukocyte rolling on activated endothelial cells.
50μg
Verified Reactivity: Human
Reported Reactivity: Chimpanzee, Cow
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Concentrated supernatant from PMA-activated human peripheral blood leukocytes
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. * Spark Blue™ 574 has a maximum excitation of 506 nm and a maximum emission of 574 nm.
Excitation Laser: Blue Laser (488 nm)
Application Notes: Additional reported applications (for the relevant formats) include: Western blotting2,3,9 and in vitro blocking of lymphocytes binding to high endothelial venules (HEV)2. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. Nos. 304853-304858).
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): Schlossman S, et al. Eds. 1995. Leucocyte Typing V. Oxford University Press. New York. Kishimoto TK, et al. 1990. Proc. Natl. Acad. Sci. USA 87:2244. (WB, Block) Jutila M, et al. 2002. J. Immunol. 169:1768. (WB) Tamassia N, et al. 2008. J. Immunol. 181:6563. (FC) PubMed Kmieciak M, et al. 2009. J. Transl. Med. 7:89. (FC) PubMed Thakral D, et al. 2008. J. Immunol. 180:7431. (FC) PubMed Charles N, et al. 2010. Nat. Med. 16:701. (FC) PubMed Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC) Koenig JM, et al. 1996. Pediatr. Res. 39:616. (WB) Shi C, et al. 2011. J. Immunol. 187:5293. (FC) PubMed Burges M, et al. 2013. Clin Cancer Res. 19:5675. PubMed Cash JL, et al. 2013. EMBO Rep. 14:999. (FC) PubMed
RRID: AB_3662508 (BioLegend Cat. No. 285168)
Structure: Selectin, single chain glycoprotein, 74-95 kD
Distribution: Majority of B cells, naïve T cells, subset of memory T and NK cells, monocytes, granulocytes, thymocytes
Function: Leukocyte homing, leukocyte tethering, rolling
Ligand/Receptor: CD34, GlyCAM, MAdCAM-1
Cell Type: B cells, Granulocytes, Monocytes, Neutrophils, NK cells, T cells, Thymocytes, Tregs
Biology Area: Cell Adhesion, Cell Biology, Costimulatory Molecules, Immunology, Innate Immunity
Molecular Family: Adhesion Molecules, CD Molecules
Antigen References: Kishimoto T, et al. 1990. P. Natl. Acad. Sci. USA 87:2244. Kishimoto T, et al. 1991. Blood 78:805.
Gene ID: NA
UniProt: View information about CD62L on UniProt.org
Clone: DREG-56
Regulatory Status: RUO
Workshop: V S056
Other Names: L-selectin, LECAM-1, LAM-1, Leu-8, TQ-1
Isotype: Mouse IgG1, κ
Q: What are Flexi-Fluors?
A: 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.
Q: How quickly will I receive my order?
A: We aim to ship Flexi-Fluors within 2-3 weeks of receipt of your order. However, depending on your location, shipping times may vary.
Q: How are Flexi-Fluors different from regular catalog products?
A: 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.
Q: How do I determine the optimal concentration for using my Flexi-Fluor? How should I titrate my antibody?
A: 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.
Q: I can’t find the antibody-dye combination that I need. When will it be available?
A: 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.
Q: I need help to validate the performance of my Flexi-Fluor. Who should I contact?
A: Please get in touch with Technical Service for assistance.
Q: Can I order more than 50 μg of a Flexi-Fluor?
A: 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.
Q: What is the expiration date of my Flexi-Fluor?
A: Expiration dates can be found on the vial label or by using our CoA lookup tool.
Q: What if my requested antibody-dye combination doesn't perform as expected?
A: Flexi-Fluor™ products have been carefully selected by our scientists for their excellent conjugation properties and broad compatibility. In the rare case that an antibody-dye combination does not perform as expected due to the unique characteristics of a specific pairing, please contact our Technical Service team. We will work with you to identify alternative options to meet your panel requirements.