Anti-Mouse EpCAM (CD326) [Clone G8.8] — Purified in vivo PLATINUM^TM Functional Grade

Clone G8.8 is a rat monoclonal antibody specific for mouse EpCAM (CD326), extensively used to identify, isolate, or characterize epithelial cells in in vivo mouse studies. Its most common applications in mouse research include flow cytometry (FACS), immunohistochemistry (IHC), immunofluorescence (IF), immunocytochemistry (ICC), and immunoprecipitation.

Key uses of clone G8.8 in in vivo mouse studies:

• Isolation and Purification: G8.8 is widely used to isolate and purify thymic epithelial cells and other EpCAM+ cell populations by flow cytometry or magnetic bead-based cell sorting.
• Tissue and Cellular Marking: Application in IHC/IF enables detection and visualization of EpCAM+ epithelial cells in tissue sections, useful in studies of tissue development, regeneration, and pathologies like cancer.
• Stem Cell and Lineage Tracing: By targeting EpCAM, G8.8 can be used in experiments tracing epithelial stem cell lineages or assessing changes in epithelial cell populations during disease or after genetic modification.
• Tumor and Developmental Studies: Since EpCAM is expressed in many epithelial tumors, the antibody is used to study tumorigenesis, tumor progression, and epithelial cell biology in mouse models.

Technical details:

• Host species: Rat
• Recognizes: Mouse EpCAM, but not human or rat EpCAM
• Common protocols: Used at concentrations of 2–5 ?g/ml for IHC/IF/ICC
• Epitope: Raised against full-length native mouse EpCAM from the TE-71 thymic epithelial cell line.

Limitations:

• The antibody does not cross-react with human or rat EpCAM, which is critical for interpretation in xenograft or humanized mouse studies.

In summary, G8.8 is a standard tool for marking, isolating, and characterizing mouse epithelial cells in vivo, especially in immunological, developmental, and cancer research.

The correct storage temperature for the sterile packaged clone G8.8 antibody is typically ?80°C upon arrival, unless otherwise specified by the manufacturer. Aliquot upon arrival and keep at ?80°C for long-term storage to preserve activity and sterility.

General guidelines for sterile antibodies (including monoclonal antibodies like G8.8):

• Aliquot and store at ?80°C for long-term storage (several months or more).
• Avoid repeated freeze-thaw cycles.
• Some antibodies may be stored at 4°C for brief periods (a few days to weeks), but this is less suitable for long-term preservation.

If the antibody is supplied lyophilized (freeze-dried) and sterile, the packaging may permit storage at 2–8°C or at room temperature until reconstitution, but most working solutions should be stored at ?80°C whenever possible for maximum stability and to retain sterility.

If you are storing the antibody in a hospital sterile supply area, ensure the temperature does not exceed 25°C for dry, unopened packaged items, but this is not appropriate for laboratory antibody reagents.

Always check the manufacturer's data sheet for clone G8.8 for specific instructions, as formulations or preservatives can affect storage recommendations. If not available, ?80°C is the safest standard for most sterile monoclonal antibody clones.

The antibody G8.8 is widely recognized as an anti-EpCAM (epithelial cell adhesion molecule) antibody. In literature, several other antibodies and proteins are commonly used together with G8.8 for identification, sorting, or functional studies involving EpCAM-expressing cells. The choice of accompanying antibodies or proteins depends on the experimental context, but the most frequently co-used markers include:

• CD44: Often co-stained with G8.8 to study cancer stem cells, as CD44 is a stem cell and adhesion marker.
• CD24: Used in combination with G8.8 for further delineation of epithelial or stem cell subpopulations, especially in cancer research.
• CD326 (EpCAM): G8.8 itself is an anti-EpCAM antibody, but other EpCAM antibodies (such as HEA125 or Ber-EP4 clones) are sometimes used for validation or in multi-color panels.
• Cytokeratins (CKs): Antibodies to cytokeratins such as CK8, CK18, or CK19 are commonly used to confirm epithelial identity in conjunction with G8.8 labeling.
• CD45: A pan-leukocyte marker used to exclude hematopoietic cells in flow cytometry, often paired with G8.8 to isolate non-hematopoietic, EpCAM-positive cells.
• Other lineage exclusion markers: Including CD31 (endothelial), CD90 (mesenchymal), and others as relevant for the system under study.

Reporter proteins and detection systems (such as enzyme-conjugates or fluorescent secondary antibodies) are technically used with G8.8 in ELISA, flow cytometry, and microscopy applications. Additionally, proteins like Protein G are utilized for antibody purification and conjugation purposes.

Example applications in literature include:

• Flow cytometry panels: G8.8 anti-EpCAM together with anti-CD44, anti-CD24, and anti-CD45 to identify and sort various cell populations.
• Immunocytochemistry/Immunohistochemistry: Co-staining with cytokeratin antibodies to validate epithelial lineage.

In studies focusing on the ABCG5/G8 transporter complex (unrelated to G8.8/EpCAM), monoclonal antibodies such as 2E10 and 11F4 have been used to form stable complexes for biochemical and structural characterization. However, this context is distinct from the G8.8 anti-EpCAM antibody.

In summary: G8.8 (anti-EpCAM) is most commonly used with antibodies against CD44, CD24, cytokeratins, CD45, and sometimes additional epithelial or lineage markers concerning isolation and characterization of epithelial and cancer stem cells.

Clone G8.8 is a rat monoclonal antibody widely used for the detection and purification of mouse EpCAM (CD326), an epithelial cell adhesion molecule important in various biological and immunological contexts. Key findings and applications from scientific literature and product documentation include:

• Species Specificity: G8.8 is specific to mouse EpCAM (CD326) and does not recognize the human or rat versions of the protein.
• Key Applications:
  • Flow Cytometry (FACS): Used to identify and sort mouse epithelial cells and subpopulations, including a small subset of CD3+CD25+EpCAM+ splenic T lymphocytes.
  • Immunohistochemistry (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC): Frequently recommended for marking epithelial cells in mouse tissues, with standard working concentrations suggested to start at 2–5?µg/mL for mouse samples.
  • Immunoprecipitation and Western Blot: Validated for these applications to detect EpCAM at ?40?kDa molecular weight in mouse samples.
• Purification of Thymic Epithelial Cells: G8.8 has been used for isolating these cells, as documented in the scientific literature (e.g., PubMed IDs 20548034 and 17127749, referenced in technical documentation).
• Epitope/Antigen Details: The immunogen for G8.8 is the full-length native EpCAM protein from the mouse TE-71 thymic epithelial cell line.
• No Cross-Reactivity: G8.8 does not cross-react with other species’ EpCAM, an important characteristic for precise mouse-specific analyses.
• Product and Repository Information: The hybridoma is available from the Developmental Studies Hybridoma Bank (DSHB), deposited by A.G. Farr (University of Washington).

Summary Table: G8.8 Clone—Key Scientific Insights

G8.8 is considered a reliable tool for mouse epithelial cell studies and immune cell profiling, particularly in thymic biology and immunology research. When referencing G8.8 in research, it is important to note its mouse specificity and acknowledge both the DSHB and the original contributor.