Alexa Fluor® 647 Rat Anti-Human Cutaneous Lymphocyte Antigen
Clone HECA-452 (RUO)
- Brand BD Pharmingen™
- Alternative Name CLA; PSGL-1; CD162; P-selectin glycoprotein ligand 1; SELPL; SELPLG
- Vol. Per Test 5 µl
- Isotype Rat WI, also known as Wistar (outbred) IgM, κ
- Reactivity Human (QC Testing)
Flow cytometry (Routinely Tested)
- Immunogen Lymphocyte-depleted Stromal Preparation of Tonsil
- Workshop No. V S075
- Storage Buffer Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The HECA-452 monoclonal antibody specifically reacts with Cutaneous Lymphocyte Associated Antigen (CLA), a carbohydrate domain shared by sialyl Lewis[x] (sLe[x]) and sialyl Lewis[a] (sLe[a]) antigens. It serves as a ligand for selectins including CD62E (E-selectin; ELAM-1). CLA is expressed on high endothelium and on lymphocytes including most T lymphocytes infiltrating cutaneous sites of inflammation. Amongst peripheral blood cells, it is expressed on monocytes and granulocytes and a subset of lymphocytes. The HECA-452 antibody is also reportedly crossreactive with the mouse CLA carbohydrate epitope that is transiently expressed by PSGL-1/CD162 on activated T cells. A number of studies suggest that CLA plays an important role in supporting leucocyte adhesive interactions and migration into extravascular tissues during inflammation.
Alexa Fluor® 647 conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of APC. However, APC tends to be brighter while Alexa Fluor® 647 is more optimal for intracellular applications. This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously.
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Preparation and Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.