CD41a Monoclonal Antibody (HIP8), NovaFluor™ Blue 510, eBioscience™

Product Specific Information

Description
The HIP8 monoclonal antibody reacts with the human CD41 molecule, the integrin alphaIIb also known as platelet GPIIb. CD41 non-covalently associates with integrin beta3 (GPIIIa, CD61) and is expressed by megakaryocytes and platelets. The CD41/CD61 complex is a receptor for fibronectin, fibrinogen, von Willebrand factor, vitronectin and thrombospondin and mediates platelets aggregation. HIP8 blocks platelet aggregation.

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

Applications Tested
This HIP8 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.6 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: 493 nm; Emission: 513 nm; Laser: 488 nm (Blue) Laser

Target Information

CD41, also known as platelet glycoprotein IIb or ITGA2B, is a protein composed of two subunits: a 120 kDa alpha subunit and a 23 kDa beta subunit. It interacts with CD61 (gpIIIa, integrin beta III) in the presence of calcium to form a functional adhesive protein receptor. Initially thought to be expressed exclusively on platelets and megakaryocytes, CD41 is also found on hematopoietic progenitors in the embryo, fetus, and adult, indicating its role in early stages of hematopoietic differentiation. CD41 plays a crucial role in platelet function and blood coagulation by mediating platelet aggregation. Upon blood vessel damage, CD41 binds to various adhesion molecules, including von Willebrand factor, fibrinogen, fibronectin, and vitronectin, facilitating platelet adhesion and aggregation. This receptor is essential for platelet function, contributing to the binding of several adhesion molecules and playing a significant role in hemostasis. Diseases associated with CD41 dysfunction include Glanzmann Thrombasthenia and Platelet type-16 bleeding disorder, highlighting its importance in normal platelet function and blood coagulation processes.