CD62L (L-Selectin) Monoclonal Antibody (MEL-14), NovaFluor™ Blue 610-30S, eBioscience™

Product Specific Information

Description
The MEL-14 monoclonal antibody reacts with mouse CD62L, a 76 kDa member of the selectin family. CD62L is expressed by neutrophils, monocytes, and subsets of T, B, and NK cells and binds a number of glycosylated, fucosylated, sulfated sialylated glycoproteins including CD34, glycam-1 and MAdCam-1. These interactions mediate rolling of lymphocytes on activated endothelium at the sites of inflammation and homing of cells to the high endothelial venules (HEV) of peripheral lymphoid tissues.

This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.

Applications Tested
The MEL-14 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.2 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.

Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.

Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.

CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (for 25 µg or 25 test products, Cat. No. C001T02F01; for 100 µg or 100 test, Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 µL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 µL of CellBlox Plus Blocking Buffer per 100 µL of cell sample (10^3 to 10^8 cells).

NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more

Excitation: 508 nm; Emission: 615 nm; Laser: 488 nm (Blue) Laser

Target Information

The human CD62L is a 74-95 kDa glycoprotein member of the selectin family of adhesion receptors. L-Selectin is comprised of an aminoterminal C-type lectin binding domain, an epidermal growth factor-like domain, two short consensus repeat (SCR) sequences homologous to those found in complement binding proteins, a short spacer region, a transmembrane region and a short cytoplasmic region. Human CD62L (L-Selectin) is constitutively expressed on all classes of leukocytes including lymphocytes (except a substantial population of memory T cells), monocytes and polymorphonuclear cells.