Detection of Human TRIM21 by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and human kidney tissue. PVDF Membrane was probed with 0.5 µg/mL of Human TRIM21 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6219) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for TRIM21 at approximately 52 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
TRIM21 in Human PBMCs.
TRIM21 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated for 8 hours with 20 ng/mL Recombinant Human IFN-gamma (Catalog # 285-IF) using Human TRIM21 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6219) 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 Non-adherent Cells.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
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.
TRIM21 (Tripartite motif-containing protein 21; also Ro(SS-A), 52 kDa Ro Protein/Ro52, and RING finger protein 81) is a 52-56 kDa member of the RING finger B box coiled coil family of proteins. It is an E3 ligase that is found in both nucleus and cytoplasm, where it is often associated with microtubules. TRIM21 ubiquitinates select proteins. In B cells, it targets the Fc fragment of misfolded IgG, providing QC on its production. In macrophages, it acts in a nondegradative manner on IRF8, promoting innate immunity. Human TRIM21 is 475 amino acids (aa) in length and contains one E3 ligase RING finger domain (aa 16-55), a B Box type zinc finger region (aa 92-123), a coiled coil region (aa 128-238) and a C-terminal SPRY/B30.2 Ig binding domain (aa 268-465). TRIM21 is reported to form trimers. Over aa 195‑293, human TRIM21 exhibits 72% aa identity with mouse TRIM21.
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