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  4. CD33 Siglec-3/CD33
  5. Siglec-3/CD33 Protein, Human (HEK293, hFc)

Siglec-3/CD33 Protein, Human (HEK293, hFc)

Cat. No.: HY-P73642
COA Handling Instructions

Siglec-3/CD33 protein is a sialic acid-binding lectin that mediates cell-cell interactions and maintains immune cell quiescence. It prefers α-2,3- and α-2,6-linked glycans with sialic acid. Siglec-3/CD33 Protein, Human (HEK293, hFc) is the recombinant human-derived Siglec-3/CD33 protein, expressed by HEK293 , with C-hFc labeled tag. The total length of Siglec-3/CD33 Protein, Human (HEK293, hFc) is 242 a.a., with molecular weight (glycosylation form) of 68-80 kDa.

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Description

Siglec-3/CD33 protein is a sialic acid-binding lectin that mediates cell-cell interactions and maintains immune cell quiescence. It prefers α-2,3- and α-2,6-linked glycans with sialic acid. Siglec-3/CD33 Protein, Human (HEK293, hFc) is the recombinant human-derived Siglec-3/CD33 protein, expressed by HEK293 , with C-hFc labeled tag. The total length of Siglec-3/CD33 Protein, Human (HEK293, hFc) is 242 a.a., with molecular weight (glycosylation form) of 68-80 kDa.

Background

The Siglec-3/CD33 protein, a sialic-acid-binding immunoglobulin-like lectin, plays a crucial role in mediating cell-cell interactions and maintaining immune cells in a resting state. It exhibits a preference for recognizing and binding alpha-2,3- and more avidly alpha-2,6-linked sialic acid-bearing glycans. Upon engagement with ligands like C1q or sialylated glycoproteins, two immunoreceptor tyrosine-based inhibitory motifs (ITIMs) within CD33's cytoplasmic tail undergo phosphorylation by Src-like kinases such as LCK. These phosphorylated ITIMs serve as docking sites for recruiting and activating protein-tyrosine phosphatases PTPN6/SHP-1 and PTPN11/SHP-2, which, in turn, regulate downstream pathways through dephosphorylation of signaling molecules. CD33's repressive effect on monocyte activation involves phosphoinositide 3-kinase/PI3K. Structurally, the protein forms homodimers through disulfide linkages and interacts with PTPN6/SHP-1 and PTPN11/SHP-2 upon phosphorylation. It also engages with C1QA via its C-terminus, activating CD33 inhibitory motifs.

Biological Activity

1.Immobilized Human Siglec-3 at 0.5 μg/mL(100 μl/well) on the plate. Dose response curve for Biotinylated Anti-Siglec-3 Antibody, hFc Tag with the EC50 of 21.8 ng/mL determined by ELISA.
2.Immobilized Human CD33 at 2 μg/mL (100 μL/well) can bind Anti-CD33 Antibody, the ED50 for this effect is 8.312 ng/mL.

  • Immobilized Human CD33 at 2 μg/mL (100 μL/well) can bind Anti-CD33 Antibody, The ED50 for this effect is 8.312 ng/mL.
Species

Human

Source

HEK293

Tag

C-hFc

Accession

P20138/AAH28152.1 (D18-H259)

Gene ID

945  [NCBI]

Molecular Construction
N-term
CD33 (D18-H259)
Accession # P20138/AAH28152.1
hFc
C-term
Synonyms
Myeloid Cell Surface Antigen CD33; Siglec-3; gp67; CD33; SIGLEC3
AA Sequence

DPNFWLQVQESVTVQEGLCVLVPCTFFHPIPYYDKNSPVHGYWFREGAIISGDSPVATNKLDQEVQEETQGRFRLLGDPSRNNCSLSIVDARRRDNGSYFFRMERGSTKYSYKSPQLSVHVTDLTHRPKILIPGTLEPGHSKNLTCSVSWACEQGTPPIFSWLSAAPTSLGPRTTHSSVLIITPRPQDHGTNLTCQVKFAGAGVTTERTIQLNVTYVPQNPTTGIFPGDGSGKQETRAGVVH

Molecular Weight

68-80 kDa (glycosylation)

Purity

Greater than 95% as determined by Tris-Bis PAGE.

Appearance

Lyophilized powder

Formulation

Lyophilized from a 0.2 μm filtered solution of PBS, pH 7.4 or 20 mM PB, 150 mM NaCl, pH 7.4.

Endotoxin Level

<1 EU/μg, determined by LAL method.

Reconstitution

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH2O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

Storage & Stability

Stored at -20°C for 2 years. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Shipping

Room temperature in continental US; may vary elsewhere.

Documentation
Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

  • Reconstitution Calculator

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The reconstitution calculator equation

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration
= ÷

The dilution calculator equation

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
× = ×
C1   V1   C2   V2

The specific activity calculator equation

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)
Unit/mg = 106 ÷ ng/mL

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Siglec-3/CD33 Protein, Human (HEK293, hFc)
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HY-P73642
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