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Your Position: 홈 > Protein > Siglec-2 > SI2-HA2H9

Alexa Fluor™ 555-Labeled Human Siglec-2 / CD22 Protein, His TagStar Staining

INTRODUCTION
QC & Assay Protocol
Citation & Background
  • Synonym
    CD22,SIGLEC2,BL-CAM,SIGLEC-2,Siglec2,SIGLEC2FLJ22814
  • Source
    Alexa Fluor 555-Labeled Human Siglec-2, His Tag (SI2-HA2H9) is produced via conjugation of AF555 to Human Siglec-2, His Tag with a new generation site-specific technology under Star Staining labeling platform. Human Siglec-2, His Tag is expressed from human 293 cells (HEK293). It contains AA Asp 20 - Arg 687 (Accession # P20273-1).
    Predicted N-terminus: Asp 20
    Request for sequence
  • Molecular Characterization
    Siglec-2 Structure

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

    The protein has a calculated MW of 92.1 kDa.

  • Conjugate

    AF555

    Excitation Wavelength: 561 nm

    Emission Wavelength: 572 nm

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

    >90% 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-2 CAR Expression
Siglec-2 CAR_T

5e5 of 293 CAR cells transfected with anti-CD22-scFv were stained with 100 μL of 10 μg/mL of Alexa Fluor 555-Labeled Human Siglec-2, His Tag (Cat. No. SI2-HA2H9) and negative control protein respectively (Fig. C and B), and non-transfected 293 cells were used as a control (Fig. A), Alexa 555 signal was used to evaluate the binding activity (QC tested).

FACS Analysis of Non-specific binding to PBMCs
Siglec-2 CAR_T

5e5 of PBMCs were stained with Alexa Fluor 555-Labeled Human Siglec-2, His Tag (Cat. No. SI2-HA2H9) 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 555 signal was used to evaluate the non-specific binding activity to PBMCs (QC tested).

  • Background
    B-cell receptor CD22 is also known as Sialic acid-binding Ig-like lectin 2 (Siglec-2), B-lymphocyte cell adhesion molecule (BL-CAM), T-cell surface antigen Leu-14, which belongs to the immunoglobulin superfamily and SIGLEC (sialic acid binding Ig-like lectin) family. CD22 mediates B-cell B-cell interactions, and may be involved in the localization of B-cells in lymphoid tissues. Siglec-2 / CD22 binds sialylated glycoproteins, one of which is CD45. Siglec2 / CD22 plays a role in positive regulation through interaction with Src family tyrosine kinases and may also act as an inhibitory receptor by recruiting cytoplasmic phosphatases via their SH2 domains that block signal transduction through dephosphorylation of signaling molecules.
  • Clinical and Translational Updates

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