TRIM21 Recombinant antibody

TRIM21 Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

240491B2

Cat no : 83530-3-RR

Synonyms

RNF 81, RING finger protein 81, EC:2.3.2.27, E3 ubiquitin-protein ligase TRIM21, 52 kDa ribonucleoprotein autoantigen Ro/SS-A

---

Validation Data Gallery

• 

  WB analysis using 83530-3-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 83530-3-RR (TRIM21 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  IHC staining of human colon cancer using 83530-3-RR

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 83530-3-RR (TRIM21 antibody) at dilution of 1:400 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of A549 using 83530-3-RR

  Immunofluorescent analysis of (4% PFA) fixed A549 cells using TRIM21 antibody (83530-3-RR, Clone: 240491B2 ) at dilution of 1:250 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) (SA00013-1), CL594-Phalloidin (red).

• 

  FC experiment of HeLa using 83530-3-RR

  1x10^6 HeLa cells were intracellularly stained with 0.25 ug TRIM21 Recombinant antibody (83530-3-RR, Clone:240491B2) and APC-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

• 

  Affinity and Kinetic Characterization of 83530-3-RR

  Biolayer interferometry (BLl) kinetic assays of 83530-3-RR against Human TRIM21 were performed. The affinity constant is 11 nM.

---

Tested Applications

• Positive WB detected in: HeLa cells, HT-29 cells, HL-60 cells, NCI-H1299
• Positive IHC detected in: human colon cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: A549 cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:200-1:800
• Immunofluorescence (IF)/ICC: IF/ICC : 1:125-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

The immunogen of 83530-3-RR is TRIM21 Fusion Protein expressed in E. coli.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: TRIM21 fusion protein Ag28377
• Full Name: tripartite motif-containing 21
• Calculated Molecular Weight: 475 aa, 54 kDa
• Observed Molecular Weight: 52 kDa
• GenBank Accession Number: BC010861
• Gene Symbol: TRIM21
• Gene ID (NCBI): 6737
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

TRIM21 has multiple N-terminal zinc finger motifs(has E3 ligase activity), a central leucine zipper, and a potential N-glycosylation site. TRIM3 is a E3 ubiquitin-protein ligase. It can form a ubiquitin ligase complex with E2 UBE2D2, which function not only for the uniquitination of USP4 and IKBKB but also for its self-ubiquitination. It has a role in the regulation of the cell cycle progression and could enhance the decapping activity of DCP2. It can exist in all mammalian cells as a ribonucleoprotein particle , which composed of a single polypeptide and 1 of 4 small RNA molecules. TRIM21 exists various isoforms and range molecular weight of isoforms is about 42-45kDa and 49-54 kDa.