A33/GPA33 Antibody (2963F) [Unconjugated]

Western blot shows lysates of COLO 205 human colorectal adenocarcinoma cells. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human A33 Monoclonal Antibody (Catalog # MAB30804) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for A33 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Colo205 cells were stained with Rabbit Anti-Human A33 Monoclonal Antibody (Catalog # MAB30804, Filled histogram) or isotype control antibody (Catalog # MAB1050, open histogram), followed by Allophycocyanin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111). View our protocol for Staining Membrane-associated Proteins.

A33 was detected in immersion fixed paraffin-embedded sections of Human Colon using Rabbit Anti-Human A33 Monoclonal Antibody (Catalog # MAB30804) at 1.7 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell membranes in epithelial cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Simple Western lane view shows lysates of human colon, loaded at 0.5 mg/ml. A specific band was detected for A33 at approximately 59 kDa (as indicated) using 20 µg/mL of Rabbit Anti-Human A33 Monoclonal Antibody (MAB30804). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

• Reactivity: Hu
• Applications: WB, Simple Western, Flow, IHC
• Clone: 2963F
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

A33/GPA33 Antibody (2963F) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: E. coli-derived recombinant human A33
  Ile22-Val235
  Accession # Q99795
• Specificity: Detects human A33 in direct ELISA.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunohistochemistry 1-15 ug/mL
  • Simple Western 20 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for A33/GPA33 Antibody (2963F) [Unconjugated]

• A33
• cell surface A33 antigen
• glycoprotein A33 (transmembrane)
• Glycoprotein A33
• GPA33
• MGC129986
• MGC129987
• transmembrane glycoprotein A33

Background

Human A33, also known as GPA33, is a 43 kDa type I transmembrane glycoprotein that belongs to the CTX (cortical thymocyte marker in Xenopus) family of cell adhesion molecules within the immunoglobulin superfamily. Other family members include CXADR, ESAM, BT-IgSF, CD2 and JAM-A-C. The extracellular domain (ECD) of human A33 is 214 amino acids (aa) in length and contains one V-type and one C2-type Ig-like domain. This ECD is 80%, 74% and 71% aa identical to canine, bovine and mouse A33 ECD, respectively. A33 is likely to be involved in cell-cell adhesion between epithelial cells.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Uniprot:
  • Q99795()