Sell Rat Monoclonal Antibody [Clone ID: MEL-14]

Specifications

Product Data
Clone Name	MEL-14
Applications	FC, FN, IHC, IP
Recommend Dilution	Flow cytometry. Immunohistochemistry. Immunoprecipitation.
Reactivity	Mouse
Host	Rat
Clonality	Monoclonal
Immunogen	Mouse B cell Lymphoma, 38C-14. Donor: Fischer Rat Spleen Fusion Partner: P3 X 63Ag8.653
Specificity	This monoclonal antibody reacts with a 90 kDa protein which is involved with the homing of lymphocytes to peripheral lymph nodes.
Formulation	PBS and 0.02% NaN3 State: Purified State: Liquid purified
Concentration	lot specific
Purification	Protein G Chromatography
Conjugation	Unconjugated
Storage Condition	Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing.
Gene Name	selectin, lymphocyte
Database Link	Entrez Gene 20343 Mouse UniProt ID P18337
Background	L-selectin is expressed on most T and B lymphocytes, neutrophils, monocytes, eosinophils.1 Pre-incubation of lymphocytes with this antibody completely and specifically blocks binding of lymphocytes to high endothelial venules (HEV) in vitro 2,3,6 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	SELL, LNHR, LYAM1, Leu-8, TQ1, gp90-MEL, LECAM1, LAM-1
Note	Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population 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 this suspension to each tube (each tube will then contain 1x10e6 cells, representing 1 test). 4. To each tube, add 0.1-0.2 µg* of this Ab. 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 (FITC Goat anti-rat IgG (H+L)) at 1:700 dilution. 9. Incubate the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes are protected from light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C in media B. 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 propidium iodide 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 (100 µl of 2M sodium azide in 100 mls). Results - Tissue Distribution: Mouse Strain: BALB/c Cell Concentration: 1x10e6 cells per test Antibody Concentration Used: 0.2 µg/10e6 cells Isotypic Control: Rat IgG2a Cell Source: Percentage of cells stained above control Thymus: 85.8% Spleen: 41.0% Lymph Node: 75.0%
Reference Data

*Delivery time may vary from web posted schedule. Occasional delays may occur due to unforeseen complexities in the preparation of your product. International customers may expect an additional 1-2 weeks in shipping.