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Invitrogen™ Cutaneous Lymphocyte Antigen (CLA) Monoclonal Antibody (HECA-452), eFluor™ 660, eBioscience™
Description
This HECA-452 monoclonal antibody reacts with human Cutaneous Lymphocyte Antigen (CLA). This cell surface glycoprotein is most highly expressed on T cells in the skin. Nevertheless, CLA can also be detected on some peripheral T and NK cells, as well as monocytes, neutrophils and CD1a+ dendritic cells. Studies have determined that this antibody recognizes sialofucosylated glycans and sialyl Lewis x (sLex). CLA, which binds E-selectin and endothelial cell-leukocyte adhesion molecule 1 (ELAM-1), is involved in T lymphocyte infiltration and adhesion at cutaneous sites of inflammation. The HECA-452 antibody has been reported to inhibit sLex binding to E-selectin; however, this property is still under debate in the literature. Applications Reported: This HECA-452 antibody has been reported for use in flow cytometric analysis, western blotting, and immunohistochemical staining of formalin-fixed paraffin embedded tissue sections. Applications Tested: This HECA-452 antibody has been tested by flow cytometric analysis on normal human peripheral blood cells. This HECA-452 antibody has also been tested by immunhistochemistry on FFPE human tissue and can be used at less than or equal to 20 μg/mL.The antibody performs optimally with low pH antigen retrieval but high pH antigen retrieval can also be used. eFluor™ 660 is a replacement for Alexa Fluor™ 647. eFluor™ 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that you...
Specifications
Specifications
| Antigen | Cutaneous Lymphocyte Antigen (CLA) |
| Applications | Flow Cytometry, Functional Assay, Immunohistochemistry (Paraffin), Western Blot, Immunocytochemistry |
| Classification | Monoclonal |
| Clone | HECA-452 |
| Concentration | 0.2 mg/mL |
| Conjugate | eFluor 660 |
| Formulation | PBS with 0.09% sodium azide; pH 7.2 |
| Gene Alias | CLA |
| Host Species | Rat |
| Purification Method | Affinity chromatography |
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Frequently Asked Questions (FAQs)
Our options will depend on the samples you are analyzing.
If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.
For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.
Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).
Yes, in-house studies have demonstrated that the eFluor 660 fluorochrome is recognized by Anti-Cy5/Alexa Fluor 647 beads. Side by side studies with Alexa Fluor 647 versus eFluor 660 conjugated antibodies have demonstrated comparable results.
The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.
The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.
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