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Your Position: Home > Protein > Siglec-3 > CD3-HA2H6

Alexa Fluor™ 647-Labeled Human Siglec-3 / CD33 Protein, His TagStar Staining

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  • Synonym
    CD33,SIGLEC3,gp67
  • Source
    Alexa Fluor 647-Labeled Human Siglec-3, His Tag (CD3-HA2H6) is produced via conjugation of AF647 to Human Siglec-3, His Tag with a new generation site-specific technology under Star Staining labeling platform. Human Siglec-3, His Tag is expressed from human 293 cells (HEK293). It contains AA Asp 18 - His 259 (Accession # AAH28152.1 ).
    Predicted N-terminus: Asp 18
  • Molecular Characterization
    Siglec-3 Structure

    This protein carries a polyhistidine tag at the C-terminus.

    The protein has a calculated MW of 43.9 kDa.

  • Conjugate

    AF647

    Excitation Wavelength: 640 nm

    Emission Wavelength: 672 nm

  • Endotoxin
    Less than 1.0 EU per μg by the LAL method.
  • Purity

    >95% as determined by SDS-PAGE.

  • Formulation

    Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 with trehalose as protectant.

    Contact us for customized product form or formulation.

  • Reconstitution

    Please see Certificate of Analysis for specific instructions.

    For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.

  • Storage

    For long term storage, the product should be stored at lyophilized state at -20°C or lower.

    Please protect from light and avoid repeated freeze-thaw cycles.

    This product is stable after storage at:

    1. -20°C to -70°C for 12 months in lyophilized state;
    2. -70°C for 3 months under sterile conditions after reconstitution.
Evaluation of CAR expression
FACS Analysis of Anti-Siglec-3 CAR Expression
 Siglec-3 CAR_T

5e5 of 293 CAR cells transfected with anti-Siglec-3-scFv were stained with 100 μL of 3 μg/mL of Alexa Fluor 647-Labeled Human Siglec-3, His Tag (Cat. No. CD3-HA2H6) and negative control protein respectively (Fig. C and B), and non- transfected 293 cells were used as a control (Fig. A), Alexa 647 signal was used to evaluate the binding activity (QC tested).

FACS Analysis of Non-specific binding to PBMCs
 Siglec-3 CAR_T

5e5 of PBMCs were stained with Alexa Fluor 647-Labeled Human Siglec-3, His Tag (Cat. No. CD3-HA2H6) and anti-CD3 antibody, washed and then analyzed with FACS. FITC signal was used to evaluate the expression of CD3+ T cells in PBMCs, and Alexa 647 signal was used to evaluate the non-specific binding activity to PBMCs (QC tested).

  • Background
    Myeloid cell surface antigen CD33 is also known as SIGLEC3, Siglecs (sialic acid binding Ig­like lectins) and GP67, is a single-pass type I membrane protein which belongs to the immunoglobulin superfamily and SIGLEC (sialic acid binding Ig-like lectin) family. Human CD33 / Siglec-3 cD encodes a 364 amino acid (aa) polypeptide with a hydrophobic signal peptide, an N-­terminal Ig-­like V­-type domain, one Ig­-like C2-­type domains, a transmembrane region and a cytoplasmic tail. CD33 / Siglec-3 usually considered myeloid-specific, but it can also be found on some lymphoid cells. In the immune response, CD33 / Siglec-3 may act as an inhibitory receptor upon ligand induced tyrosine phosphorylation by recruiting cytoplasmic phosphatase(s) via their SH2 domain(s) that block signal transduction through dephosphorylation of signaling molecules. CD33 / Siglec-3 induces apoptosis in acute myeloid leukemia.
  • Clinical and Translational Updates

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