ANTI-TRIM25 CLONE 1B8 ZOOMAB« RABBIT MONOCLONAL (C15-1669-389)

General description

ZooMAb^® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb^® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb^® antibodies are reliably available and ready to ship when you need them.

Specificity

Clone 1B8 is a ZooMAb^® rabbit recombinant monoclonal antibody that specifically detects TRIM25. It targets an epitope within the internal region.

Immunogen

His-tagged recombinant fragment corresponding to 236 amino acids from the internal region of human TRIM25.

Application

Quality Control Testing

Evaluated by Western Blotting in HepG2 cell lysate.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected TRIM25 in HepG2 cell lysate.

Tested Applications

Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected TRIM25 in HeLa cell lysate.

Immunohistochemistry (Paraffin) Analysis: A 1:100-1,000 dilution from a representative lot detected TRIM25 in Human pancreas and Human liver tissue sections.

Affinity Binding Assay A representative lot of this antibody bound recombinant fragment of TRIM25 protein with a KD of 3.3 x 10-8 in an affinity binding assay.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Target description

E3 ubiquitin/ISG15 ligase TRIM25 (UniProt: Q14258; also known as EC: 2.3.2.27, Estrogen-responsive finger protein, RING finger protein 147, RING-type E3 ubiquitin transferase, RING-type E3 ubiquitin transferase, TRIM25, Tripartite motif-containing protein 25, Ubiquitin/ISG15-conjugating enzyme TRIM25, Zinc finger protein 147) is encoded by the TRIM25 (also known as EFP, RNF147, ZNF147) gene (Gene ID: 7706) in human. TRIM25 is a ubiquitously expressed, homodimeric, E3 ubiquitin ligase that is involved in various cellular processes, including regulation of the innate immune response against viruses. TRIM25-mediated ubiquitination of the cytosolic pattern recognition receptor RIG-I is an essential step for initiation of the intracellular antiviral response. Recognition of viral RNA by RIG-I is reported to expose its 2 CARD domains for binding to the C-terminal SPRY domain of TRIM25. The RING E3 ligase activity of TRIM25 then conjugates K63-polyubiquitin chains to residues various lysine residues in RIG-I. The K63-linked ubiquitin chains on RIG-I promote its interaction with mitochondrial antiviral signaling protein (MAVS) and subsequent downstream activation of intracellular antiviral signaling. TRIM25 many also negatively regulate RIG-I through stabilization of the ubiquitin-like protein FAT10 that binds to RIG-I in a non-covalent manner and sequesters it from the signaling platform inhibiting interferon regulatory factor 3  (IRF3) and NF-κB activation. This process limits the inflammatory response and reduces host damage. TRIM25 also plays an essential role in the antiviral activity of ZAP that inhibits the replication of certain viruses.  It mediates lysine 63-linked polyubiquitination of ZAP, which is required for its optimal binding to target mRNA. TRIM25 contains a RING domain, two B-boxes domains, a CC dimerization domain, and a C-terminal SPRY domain. Its RING-type zinc finger (aa 13-54) is reported to be important for ISG15 E3 ligase activity and autoISGylation. This ZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, Affinity^™, reproducibility, and stability over conventional monoclonals. (Ref.: Yang, E, et al. (2022). PLoS Pathog. 18(9); e1010743; María Martín-Vicente, M., et al. (2017). Front. Immunol. 8; 1187; Nguyen, NTH., et al. (2016). Sci. Rep. 6; 23377).

Physical form

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.

Reconstitution

300 µg/mL after reconstitution at 25 µL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

Legal Information

Affinity is a trademark of Mine Safety Appliances Co.

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.