TRIM25 Recombinant Rabbit mAb (S-4069-59)

Catalog Number: S0B60517 Application: WB, IHC-P, ICC Reactivity: Hu, Mk Conjugation: Unconjugated Brand: Starter

Price:

$100 USD

Size: 25μl 100μl

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Product Details

Product Specification

Host	Rabbit
Antigen	TRIM25
Immunogen	Synthetic Peptide
Location	Cytoplasm
Accession	Q14258
Clone Number	S-4069-59
Antibody Type	Recombinant mAb
Isotype	IgG
Application	WB, IHC-P, ICC
Reactivity	Hu, Mk
Positive Sample	HeLa, K-562, HepG2, COLO 205, COS-7
Purification	Protein A
Concentration	2 mg/ml
Conjugation	Unconjugated
Physical Appearance	Liquid
Storage Buffer	PBS, 40% Glycerol, 0.05% BSA, 0.02% sodium azide
Stability & Storage	12 months from date of receipt / reconstitution, -20 °C as supplied

Dilution

application	dilution	species
WB	1:1000-1:2000	Hu, Mk
IHC-P	1:400	Hu
ICC	1:200	Hu

Background

TRIM25, is an important E3 ubiquitin ligase that plays a critical role in innate immunity and tumorigenesis. Its classic function is as a core regulatory factor of the RIG-I-mediated antiviral signaling pathway, activating the type I interferon response by catalyzing K63-linked polyubiquitination of RIG-I. Recent structural studies have revealed the molecular mechanism of the interaction between the PRYSPRY domain of TRIM25 and the 2CARD domain of RIG-I, and this binding promotes RIG-I dimerization and further tetramerization, thereby activating downstream signaling. In embryonic stem cells, TRIM25 also exhibits an antiviral function independent of the interferon pathway, translocating from the nucleus to the cytoplasm upon viral infection, directly binding viral RNA, and forming condensates with G3BP1 to restrict viral replication. Furthermore, TRIM25 participates in the regulation of multiple cell death pathways, including apoptosis, pyroptosis, necroptosis, ferroptosis, and autophagy, and its abnormal expression is associated with chemotherapy resistance and poor prognosis in various tumors. In non-small cell lung cancer, acetylation modification of TRIM25 can enhance its protein stability, thereby driving tumor progression by promoting epithelial-mesenchymal transition and metastasis. In glioblastoma, TRIM25 regulates the PRMT1/c-MYC pathway by targeting the splicing factor NONO, promoting tumor cell growth and invasion.

Picture

Western Blot

WB result of TRIM25 Recombinant Rabbit mAb
Primary antibody: TRIM25 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: K-562 whole cell lysate 20 µg
Lane 3: HepG2 whole cell lysate 20 µg
Lane 4: COLO 205 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 71 kDa
Observed MW: 68 kDa

WB result of TRIM25 Recombinant Rabbit mAb
Primary antibody: TRIM25 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: COS-7 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 71 kDa
Observed MW: 60 kDa

Immunohistochemistry

IHC shows positive staining in paraffin-embedded human kidney. Anti-TRIM25 antibody was used at 1/400 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human spleen. Anti-TRIM25 antibody was used at 1/400 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human breast cancer. Anti-TRIM25 antibody was used at 1/400 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human ovarian cancer. Anti-TRIM25 antibody was used at 1/400 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

Immunocytochemistry

ICC shows positive staining in HeLa cells. Anti-TRIM25 antibody was used at 1/200 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).