CD33 Human, Sf9

CD33 Human Recombinant, Sf9
Cat. No.
BT28391
Source
Sf9, Baculovirus cells.
Synonyms

Myeloid cell surface antigen CD33 isoform 1, CD33, FLJ00391, p67, SIGLEC-3, SIGLEC3, gp67.

Appearance
Sterile Filtered colorless solution.
Purity

Greater than 90.0% as determined by SDS-PAGE.

Usage
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description

CD33 produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain (18-259 a.a.) and fused to a 239 aa hIgG-His Tag at C-terminus containing a total of 484 amino acids and having a molecular mass of 54kDa. CD33 shows multiple bands between 50-70kDa on SDS-PAGE, reducing conditions and purified by proprietary chromatographic techniques. 

Product Specs

Introduction
CD33, also known as Siglec-3, is a transmembrane receptor expressed on myeloid progenitor cells, mature monocytes, macrophages, and microglial cells. It plays a role in cell adhesion and signaling within the immune system. CD33 recognizes and binds to sialic acid residues on glycans, with a preference for alpha-2,6-linked sialic acid. This interaction can mediate cell-cell adhesion. CD33 also functions as an inhibitory receptor. Upon ligand binding, CD33 becomes tyrosine phosphorylated, leading to the recruitment of cytoplasmic phosphatases. These phosphatases then dephosphorylate downstream signaling molecules, effectively blocking signal transduction pathways. Dysregulation of CD33 function has been implicated in various diseases, including cancer and neurodegenerative disorders. In acute myeloid leukemia (AML), for instance, CD33 is often overexpressed and contributes to disease progression. Notably, CD33 is a therapeutic target in AML, with antibody-drug conjugates like gemtuzumab ozogamicin used to specifically target and eliminate CD33-positive leukemia cells.
Description
CD33, a single-pass type I transmembrane protein, is produced in Sf9 insect cells using baculovirus expression system. This recombinant protein encompasses amino acids 18 to 259 of the extracellular domain of human CD33, fused to a C-terminal His-tag and a human IgG-Fc domain. The resulting protein has a total of 484 amino acids and a molecular weight of approximately 54 kDa. SDS-PAGE analysis under reducing conditions reveals multiple bands within the 50-70 kDa range, likely representing glycosylated forms of the protein. The purification process involves proprietary chromatographic methods to ensure high purity.
Physical Appearance
Clear, colorless solution, sterile-filtered.
Formulation
The CD33 protein is supplied at a concentration of 0.25 mg/ml in a buffer consisting of phosphate-buffered saline (PBS) at pH 7.4 and 10% glycerol.
Stability
For short-term storage (up to 4 weeks), the CD33 protein should be kept refrigerated at 4°C. For long-term storage, it is recommended to freeze the protein at -20°C. To prevent protein degradation during freezing and thawing, it is advisable to add a carrier protein such as albumin (HSA or BSA) at a concentration of 0.1%. Repeated freeze-thaw cycles should be avoided to maintain protein integrity and activity.
Purity
The purity of the CD33 protein is greater than 90%, as determined by SDS-PAGE analysis.
Synonyms

Myeloid cell surface antigen CD33 isoform 1, CD33, FLJ00391, p67, SIGLEC-3, SIGLEC3, gp67.

Source
Sf9, Baculovirus cells.
Amino Acid Sequence

ADLDPNFWLQ VQESVTVQEG LCVLVPCTFF HPIPYYDKNS PVHGYWFREG AIISGDSPVA TNKLDQEVQE ETQGRFRLLG DPSRNNCSLS IVDARRRDNG SYFFRMERGS TKYSYKSPQL SVHVTDLTHR PKILIPGTLE PGHSKNLTCS VSWACEQGTP PIFSWLSAAP TSLGPRTTHS SVLIITPRPQ DHGTNLTCQV KFAGAGVTTE RTIQLNVTYV PQNPTTGIFP GDGSGKQETR AGVVHLEPKS CDKTHTCPPC PAPELLGGPS VFLFPPKPKD TLMISRTPEV TCVVVDVSHE DPEVKFNWYV DGVEVHNAKT KPREEQYNST YRVVSVLTVL HQDWLNGKEY KCKVSNKALP APIEKTISKA KGQPREPQVY TLPPSRDELT KNQVSLTCLV KGFYPSDIAV EWESNGQPEN NYKTTPPVLD SDGSFFLYSK LTVDKSRWQQ GNVFSCSVMH EALHNHYTQK SLSLSPGKHH HHHH.

Product Science Overview

Structure and Expression

CD33 is a transmembrane protein predominantly expressed on the surface of myeloid cells, including monocytes, macrophages, and myeloid progenitor cells. The protein consists of an extracellular domain that binds to sialic acids, a single transmembrane region, and an intracellular domain that contains immunoreceptor tyrosine-based inhibitory motifs (ITIMs). These ITIMs are essential for the inhibitory signaling functions of CD33.

Function

CD33 functions as an inhibitory receptor in the immune system. Upon binding to its ligands, which are typically sialic acid-containing glycoproteins, CD33 undergoes tyrosine phosphorylation. This phosphorylation recruits cytoplasmic phosphatases, such as SHP-1 and SHP-2, which dephosphorylate signaling molecules and inhibit cellular activation. This mechanism helps regulate immune responses and maintain immune homeostasis.

In addition to its role in immune regulation, CD33 has been implicated in the pathogenesis of acute myeloid leukemia (AML). CD33 is expressed on the surface of leukemic blasts in most AML patients, making it a target for therapeutic interventions. Antibody-drug conjugates targeting CD33, such as gemtuzumab ozogamicin, have been developed for the treatment of AML.

Recombinant CD33 (Human, Sf9)

Recombinant CD33 protein is produced using the baculovirus expression system in Sf9 insect cells. This system allows for the production of glycosylated proteins that closely resemble their native forms. The recombinant CD33 protein typically consists of the extracellular domain of CD33 fused to a tag, such as a His-tag, to facilitate purification and detection.

The recombinant CD33 protein produced in Sf9 cells is a single, glycosylated polypeptide chain with a molecular mass of approximately 54 kDa . It is purified using chromatographic techniques to achieve high purity and is often used in research applications to study the structure, function, and interactions of CD33.

Applications

Recombinant CD33 protein is widely used in various research applications, including:

  • Structural studies: Understanding the three-dimensional structure of CD33 and its interactions with ligands.
  • Functional assays: Investigating the signaling pathways and inhibitory functions of CD33.
  • Drug development: Screening and characterizing potential therapeutic agents targeting CD33 for the treatment of AML and other diseases.

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