Human/Mouse TRIM32 Antibody

  
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human and mouse TRIM32 in Western blots.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human TRIM32
    Arg105-Lys204
    Accession # Q13049
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.5 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
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.
Immunocytochemistry
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.
Immunohistochemistry
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
  • Reconstitution
    Sterile PBS to a final concentration of 0.2 mg/mL.
  • Shipping
    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.
Background: TRIM32
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.
  • Long Name:
    Tripartite Motif-containing Protein 32
  • Entrez Gene IDs:
    22954 (Human); 69807 (Mouse); 313264 (Rat)
  • Alternate Names:
    72 kDa Tat-interacting protein; BBS11; BBS1172-KD; EC 6.3.2.-; HT2A; HT2Alimb girdle muscular dystrophy 2H (autosomal recessive); LGMD2H; TATIP; TRIM32; tripartite motif containing 32; Zinc finger protein HT2A; zinc-finger protein HT2A
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