L-Selectin antibody (Biotin)

Catalog No. ABIN114232

Product Details anti-L-Selectin Antibody (Biotin)

Target: L-Selectin (SELL) (Selectin L (SELL))

Reactivity: Mouse

Host: Rat

Clonality: Monoclonal

Conjugate: This L-Selectin antibody is conjugated to Biotin

Application: Flow Cytometry (FACS)

Specificity: This monoclonal antibody reacts with a 90 kDa protein which is involved with the homing of lymphocytes to peripheral lymph nodes.

Purification: Protein G Chromatography

Immunogen: Mouse B cell Lymphoma, 38C-14 Donor: Fischer Rat Spleen Fusion Partner: P3 X 63Ag8.653

Clone: MEL-14

Isotype: IgG2a

Application Details

Application Notes: Flow cytometry. Immunohistochemistry.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation with Lympholyte®-M cell separation medium. 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 µl of thissuspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test). 4. To each tube, add 0. 1-0. 2 µg* of this Ab per 10e6 cells. 5. Vortex the tubes to ensure thorough mixing of antibody and cells. 6. Incubate the tubes for 30 minutes at 4°C. 7. Wash 2 times at 4°C. 8. Add 100 µl of secondary antibody (Streptavidin-FITC) at a 1: 500 dilution. 9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protectedfrom light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C. 11. Resuspend the cell pellet in 50 µl ice cold media B. 12. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidiumiodide at 0. 5 mg/ml in PBS. This stains dead cells by intercalating in DNA. Media: A. Phosphate buffered saline (pH 7. 2) + 5% normal serum of host species + sodium azide(100 µl of 2M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7. 2) + 0. 5% Bovine serum albumin + sodium azide (100µl of 2M sodium azide in 100 mls). Results - Tissue Distribution: Mouse Strain: BALB/cCell Concentration: 1x10e6 cells per testAntibody Concentration Used: 0. 2 µg/10e6 cellsIsotypic Control: Biotin Rat IgG2aCell Source: Percentage of cells stained above controlThymus: 59. 8%Spleen: 66. 3%Lymph Node: 75. 8%

Restrictions: For Research Use only

Handling

Concentration: 0.1 mg/mL

Buffer: PBS, 0.02 % NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/mL

Preservative: Sodium azide

Precaution of Use: This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeated freezing and thawing.

Storage: 4 °C/-20 °C

Storage Comment: Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.

Target Details for L-Selectin

Target: L-Selectin (SELL) (Selectin L (SELL))

Alternative Name: CD62L / L-Selectin (SELL Products)

Target Type: Chemical

Background: L-selectin is expressed on most T and B lymphocytes, neutrophils, monocytes and eosinophils.1 Pre-incubation of lymphocytes with this antibody completely and specifically blocks binding of lymphocytes to high endothelial venules (HEV) in vitro2,3,6 and the migration of lymphocytes to lymph nodes in vivo.2,3 Polymorphonuclear cells preincubated with this antibody do not migrate to the inflammatory foci.Synonyms: CD62 antigen-like family member L, LAM-1, LECAM1, LNHR, LYAM1, Leu-8, Leukocyte adhesion molecule 1, Leukocyte surface antigen Leu-8, Leukocyte-endothelial cell adhesion molecule 1, Lymph node homing receptor, SELL, TQ1, gp90-MEL

Gene ID: 20343

NCBI Accession: NP_035476

UniProt: P18337