TRIM65 Antibody (Center)

Affinity Purified Rabbit Polyclonal Antibody (Pab)

All lanes: Anti-TRIM65 Antibody (Center) at 1:1000 dilution Lane 1: Hela whole cell lysate Lane 2: U-251 MG whole cell lysate Lane 3: 293T whole cell lysate Lane 4: NCI-H460 whole cell lysate Lane 5: Mouse testis lysate Lane 6: Mouse brain lysate Lysates/proteins at 20 µg per lane. Secondary: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ASP1615) at 1/15000 dilution. Observed band size: 65 KDa Blocking/Dilution buffer: 5% NFDM/TBST.
TRIM65 Antibody (Center) (Cat. #AP11814c)immunohistochemistry analysis in formalin fixed and paraffin embedded human prostate carcinoma followed by peroxidase conjugation of the secondary antibody and DAB staining.This data demonstrates the use of TRIM65 Antibody (Center) for immunohistochemistry. Clinical relevance has not been evaluated.
Confocal immunofluorescent analysis of TRIM65 Antibody (Center)(Cat#AP11814c) with NCI-H460 cell followed by Alexa Fluor 488-conjugated goat anti-rabbit lgG (green). Actin filaments have been labeled with Alexa Fluor 555 phalloidin (red).DAPI was used to stain the cell nuclear (blue).
TRIM65 Antibody (Center) (Cat. #AP11814c) flow cytometric analysis of NCI-H460 cells (right histogram) compared to a negative control cell (left histogram).FITC-conjugated goat-anti-rabbit secondary antibodies were used for the analysis.

Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC-P, IF, FC, E
Primary Accession	Q6PJ69
Other Accession	NP_775818.2
Reactivity	Human
Host	Rabbit
Clonality	Polyclonal
Isotype	Rabbit IgG
Calculated MW	57353 Da
Antigen Region	327-355 aa

Additional Information
Gene ID	201292
Other Names	Tripartite motif-containing protein 65, TRIM65
Target/Specificity	This TRIM65 antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 327-355 amino acids from the Central region of human TRIM65.
Dilution	WB~~1:1000 IHC-P~~1:100~500 IF~~1:10~50 FC~~1:10~50 E~~Use at an assay dependent concentration.
Format	Purified polyclonal antibody supplied in PBS with 0.05% (V/V) Proclin 300. This antibody is prepared by Saturated Ammonium Sulfate (SAS) precipitation followed by dialysis against PBS.
Storage	Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Precautions	TRIM65 Antibody (Center) is for research use only and not for use in diagnostic or therapeutic procedures.

Protein Information
Name	TRIM65
Function	E3 ubiquitin ligase that plays a role in several processes including innate immnity, autophagy or inflammation (PubMed:28594402, PubMed:34512673). Negatively regulates miRNAs by modulating the ubiquitination and stability of TNRC6A, a protein involved in RNA- mediated gene silencing by both micro-RNAs (miRNAs) and short interfering RNAs (PubMed:24778252). This ubiquitination results in the suppressed expression of miR-138-5p leading to increased autophagy (PubMed:31160576). Upon enteroviral infection, promotes 'Lys-63'- mediated ubiquitination activation of IFIH1/MDA5 leading to innate signaling cascade (PubMed:28594402). Mechanistically, selectively recognizes MDA5 filaments that occur on dsRNAs (PubMed:33373584). Plays also a role in limitation of inflammation through different mechanisms. First, promotes 'Lys-48'-mediated ubiquitination of VCAM1 leading to its degradation and limitation of LPS-induced lung inflammation (PubMed:31310649). In addition, negatively regulates inflammasome activation by promoting 'lys48'-linked ubiquitination of NLRP3 which is critical for the inhibition of NLRP3 inflammasome activation in resting macrophages (PubMed:34512673).
Cellular Location	Cytoplasm