ChREBP (carbohydrate response element-binding protein) is a glucose responsive basic helix-loop-helix/leucine zipper (bHLH/LZ) transcription factor that binds MLX and then carbohydrate response element /ChoRE for the induction of genes involved in glycolysis, de novo lipogenesis (DNL), and fatty acid desaturation. ChREBP’s target genes includes glucokinase (GCK), fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), pyruvate kinase/liver type pyruvate kinase (PK1/ PKLR), delta-9-desaturase (SCD/SCD1) etc. ChREBP is expressed in a number of mammalian tissues such as liver, skeletal muscle, white/brown adipose, heart, kidney, cerebellum and intestine. Increased hepatic ChREBP expression has been suggested to result in development of non-alcoholic fatty liver disease (NAFLD) and obesity through the conversion of carbohydrates into triglycerides. Recent findings have suggested that besides glucose-lipids homeostasis, it implicates in pathways linked to insulin signaling and tumorigenesis/cancers also (1).
Novus Biologicals offers several ChREBP Antibodies (including a newly launched mAB CHREBP Antibody clone 2D9NB) and these antibodies are available in unconjugated as well as in multiple pre-conjugated formats. Our ChREBP antibodies are cited in many publications from journals of high repute.
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ICC/IF with CHREBP Antibody [NB400-135] HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-ChREBP (NB400-135) at a 1:200 dilution overnight at 4C and detected with an anti-rabbit Dylight 488 at a 1:500 dilution (Green; see cytoplasmic-nuclear staining of ChREBP). Anti-alpha tubulin was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red). Nuclei were counterstained with DAPI (Blue). |
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IHC-P with CHREBP Antibody (2D9NB) [NBP2-44307] - IHC analysis of a formalin fixed and paraffin embedded tissue section of human hepatocellular carcinoma using 1:200 dilution of CHREBP antibody (clone 2D9NB) with HRP-DAB detection and hematoxylin counterstaining. The antibody generated a diffused to punctate staining pattern in the cytoplasm of the cancer cells. Some cells depicted nuclear positivity also while the stroma was largely negative. |
Levin's team from University of California used Novus' CHREBP Antibody (NB400-136) in Western blot/WB as well as confocal staining (Immunocytochemistry/Immunofluorescence; ICC/IF) assays and showed that the membrane estrogen receptor alpha inhibits nuclear translocation of CHREBP in insulin treated differentiated 3T3-L1 cells (AMPK dependent effect) (2). In another study, the authors employed ChREBP Antibody [NB400-135] for WB analysis of liver and white adipose tissue/WAT from transgenic mice expressing constitutively active isoform of ChREBP (caChREBP), and showed that binding protein 4/FABP4-Cre-mediated expression of caChREBP results in an increased expression of the lipogenic ChREBP target genes in WAT and protected against adiposity, insulin sensitivity and hepatosteatosis (3). In an article published in The International Journal of Biochemistry & Cell Biology, the authors used Novus ChREBP antibody [NB400-135] for WB and ICC/IF assays in HCT116 colorectal cancer and HepG2 hepatocellular carcinoma cells. They showed that ChREBP co-localize to nuclei/cytoplasm of cells with flightless I homolog (FLII), and that FLII acts as a component of the ChREBP transcriptional complex and negatively regulates ChREBP activity in cancer cells (4).
Compiled By: Subhash Gangar
References
ChREBP Products from Novus Biologicals