CD43 Monoclonal Antibody (eBio84-3C1 (84-3C1)), NovaFluor™ Blue 690, eBioscience™

Product Specific Information

Description
The monoclonal antibody ebio84-3C1 reacts with CD43, which is also known as leukosialin, galactoglycoprotein and sialophorin.

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

Applications Tested
This ebio84-3C1 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 µL (0.4 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.

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: 494 nm; Emission: 688 nm; Laser: 488 nm (Blue) Laser

Target Information

CD43, also known as leukosialin or sialophorin, is a transmembrane mucin-like protein characterized by a high negative charge. It is expressed on the surface of most hematopoietic cells, excluding resting B lymphocytes. CD43 serves both adhesive and anti-adhesive functions, contributing to a repulsive barrier that can interfere with cellular adhesion while also promoting leukocyte aggregation in certain contexts. CD43 is involved in the activation of T cells, B cells, NK cells, and monocytes. Its expression decreases upon activation due to proteolytic processing. The counter-receptor for CD43 is CD169/SIGLEC-1, which is expressed on macrophages. CD43 interacts with actin-binding proteins ezrin and moesin, playing a regulatory role in remodeling T-cell morphology and regulating cell-cell interactions during lymphocyte traffic. It enhances LFA-1 adhesiveness while counteracting LFA-1 induction via other receptors. CD43 signaling induces the functionally active tumor suppressor p53 protein, but in cases of p53 and ARF deficiency, it can promote tumor proliferation and viability. CD43 is an important modulator of leukocyte functions, with dysfunctions associated with diseases such as Wiscott-Aldrich Syndrome and Adenoid Basal Cell Carcinoma.