Detection of Human and Mouse TRIM32 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, PC‑3 human prostate cancer cell line, and A20 mouse B cell lymphoma cell line. PVDF Membrane was probed with 0.5 µg/mL of Sheep Anti-Human/Mouse TRIM32 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6515) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for TRIM32 at approximately 72 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
TRIM32 in HeLa Human Cell Line. TRIM32 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human/Mouse TRIM32 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6515) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
TRIM32 in Human Prostate Cancer Tissue. TRIM32 was detected in immersion fixed paraffin-embedded sections of human prostate cancer tissue using Sheep Anti-Human/Mouse TRIM32 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6515) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & 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.
TRIM32 (Tripartite motif-containing protein 32; also 72 kDa TAT-interacting protein, and zinc finger protein HT2A) is a 72-82 kDa member of the TRIM/RBCC family of proteins. It is an E3 ligase that is found in structures termed nuclear and cytoplasmic bodies. Cells known to express TRIM32 are diverse and include fibroblasts, keratinocytes, skeletal muscle cells and neurons. TRIM32 ubiquitinates select proteins such as c-myc, Abi2, actin and dysbindin. Human TRIM32 is 653 amino acids (aa) in length. It contains one E3 ligase RING finger domain (aa 20-65), a B-Box type zinc-finger region (aa 103-133), a coiled-coil region (aa 138-197), five NHL repeats (aa 358-646) and three utilized phosphorylation sites (Ser328/335/339). TRIM32 has the potential to form homomultimers. Over aa 105-204, human TRIM32 exhibits 95% aa identity with mouse TRIM32.
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