CHREBP Antibody

Images

 
Western Blot: CHREBP Antibody [NB400-135] - Detection of ChREBP in liver nuclear extracts from well-fed rats. 7% SDS gel, 1:1,000 dilution of NB400-135. Photo courtesy of Dr. Uyeda, UT Southwestern University.
Immunocytochemistry/ Immunofluorescence: 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 ...read more
Immunohistochemistry: CHREBP Antibody [NB400-135] - Colon, Epithelium 40X.
Western Blot: CHREBP Antibody [NB400-135] - Detection of ChREBP in 20 ug of human hepatocyte lysate using NB400-135. 5-10 second film exposure.
Immunocytochemistry/ Immunofluorescence: CHREBP Antibody [NB400-135] - Immunofluorescent staining of a human hepatocyte using NB400-135. Overlay staining of ChREBP (rhodamine red) and DAPI nuclear staining (blue).
Immunohistochemistry: CHREBP Antibody [NB400-135] - Analysis of CHREBP in mouse liver using DAB with hematoxylin counterstain.

Product Details

Summary
Reactivity Hu, Mu, RtSpecies Glossary
Applications WB, ChIP, IB, ICC/IF, IHC, IHC-P, IP
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1 mg/ml

CHREBP Antibody Summary

Immunogen
A C-terminal synthetic peptide made to the human CHREBP protein sequence (between residues 800-852). [UniProt# Q9NP71, Isoform 1/Alpha]
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
MLXIPL
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:1000
  • Chromatin Immunoprecipitation 1:10-1:500
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:100-1:500
  • Immunohistochemistry 1:100
  • Immunohistochemistry-Paraffin 1:100
  • Immunoprecipitation 1:10-1:500
Application Notes
This CHREBP antibody is useful in ChIP (PMID: 21282101), Immunohistochemistry paraffin embedded sections, Immunocytochemistry/Immunofluorescence and Western blot, where a band is seen at ~95 kDa. Use in immunoblotting reported in scientific literature (PMID 26181104) The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Theoretical MW
95 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Control Peptide
CHREBP Peptide (NB400-135PEP)
Reviewed Applications
Read 5 Reviews rated 4.2
using
NB400-135 in the following applications:

Publications
Read Publications using
NB400-135 in the following applications:

  • 21 publications
  • 1 publication
  • 3 publications
  • 1 publication
  • IP
    4 publications
  • WB
    43 publications

Reactivity Notes

Human, rat and mouse (PMID 21835137).

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.02% Sodium Azide
Concentration
1 mg/ml
Purity
Immunogen affinity purified

Alternate Names for CHREBP Antibody

  • BHLHD14
  • CHREBPcarbohydrate-responsive element-binding protein
  • Class D basic helix-loop-helix protein 14
  • MIOWS basic-helix-loop-helix leucine zipper protein
  • MLX interacting protein-like
  • MLX interactor
  • MLX-interacting protein-like
  • MONDOBWilliams-Beuren syndrome chromosomal region 14 protein
  • WBSCR14Williams-Beuren syndrome chromosome region 14 protein 1
  • Williams Beuren syndrome chromosome region 14
  • Williams-Beuren syndrome chromosome region 14 protein 2
  • WS-bHLHbHLHd14carbohydrate response element binding protein

Background

Carbohydrate response element-binding protein (ChREBP) is a glucose sensitive basic helix-loop-helix/leucine zipper transcription factor which heterodimerise with MLX and binds to carbohydrate response element (ChoRE) on DNA for the induction genes involved in glycolysis, de novo lipogenesis (DNL), and fatty acid desaturation. Some of the target genes of ChREBP are 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 as alpha and beta alternatively spliced isoforms, and among these two isoforms, the alpha-isoform is activated in a glucose-dependent manner. ChREBP is expressed in the cytoplasm and nuclei of a number of mammalian tissues, with high levels found in liver, skeletal muscle, white/brown adipose, heart, kidney, cerebellum and intestine. Glucose stimulates ChREBP transcriptional expression and activates the transactivity of ChREBP protein. It is activated independent of insulin and recent findings have suggested that together with its binding partner Mlx, ChREBP implicates in metabolic processes, insulin signaling related pathways, and also tumorigenesis/cancers. Also, 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 (de novo lipogenesis).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Publications for CHREBP Antibody (NB400-135)(88)

We have publications tested in 3 confirmed species: Human, Mouse, Rat.

We have publications tested in 6 applications: ChIP, EMSA, ICC/IF, IHC-P, IP, WB.


Filter By Application
ChIP
(21)
EMSA
(1)
ICC/IF
(3)
IHC-P
(1)
IP
(4)
WB
(43)
All Applications
Filter By Species
Human
(14)
Mouse
(35)
Rat
(17)
All Species
Showing Publications 1 - 10 of 88. Show All 88 Publications.
Publications using NB400-135 Applications Species
Kim YS, Kim M, Choi MY et al. Aralia elata (Miq) Seem Extract Decreases O-GlcNAc Transferase Expression and Retinal Cell Death in Diabetic Mice. J Med Food. 2017 Oct 01 [PMID: 29040017] (WB, Mouse) WB Mouse
Iroz A, Montagner A, Benhamed F et al. A Specific ChREBP and PPARa Cross-Talk Is Required for the Glucose-Mediated FGF21 Response. Cell Rep. 2017 Oct 10 [PMID: 29020627] (ChIP, Mouse) ChIP Mouse
Softic S, Gupta MK, Wang GX et al. Divergent effects of glucose and fructose on hepatic lipogenesis and insulin signaling. J. Clin. Invest. 2017 Nov 01 [PMID: 28972537] (WB, Mouse) WB Mouse
Heidenreich S, Witte N, Weber P et al. Retinol saturase coordinates liver metabolism by regulating ChREBP activity Nat Commun 2017 Aug 30 [PMID: 28855500] (WB, Mouse) WB Mouse
Kim YS, Kim M, Choi MY et al. Metformin protects against retinal cell death in diabetic mice Biochem. Biophys. Res. Commun. 2017 Aug 23 [PMID: 28843855] (Mouse) Mouse
Fan Q, Norgaard RC, Bindesboll C et al. LXRalpha Regulates Hepatic ChREBPalpha Activity and Lipogenesis upon Glucose, but Not Fructose Feeding in Mice Nutrients 2017 Jun 29 [PMID: 28661453] (WB, Mouse) WB Mouse
Cho JH, Kim GY, Pan CJ et al. Downregulation of SIRT1 signaling underlies hepatic autophagy impairment in glycogen storage disease type Ia. PLoS Genet. 2017 May 01 [PMID: 28558013] (IHC-P, Mouse) IHC-P Mouse
Janssens S, Ciapaite J, Wolters JC et al. An In Vivo Magnetic Resonance Spectroscopy Study of the Effects of Caloric and Non-Caloric Sweeteners on Liver Lipid Metabolism in Rats. Nutrients. 2017 May 10 [PMID: 28489050] (WB, Rat) WB Rat
Na TY, Kim GH, Oh HJ et al. The trisaccharide raffinose modulates epidermal differentiation through activation of liver X receptor. Sci Rep. Mar 7 2017 [PMID: 28266648] (ChIP, Human) ChIP Human
Al-Oanzi ZH, Fountana S, Moonira T et al. Opposite effects of a glucokinase activator and metformin on glucose-regulated gene expression in hepatocytes. Diabetes Obes Metab. 2017 Feb 16 [PMID: 28206714] (ChIP, Rat) ChIP Rat
Show All 88 Publications.

Reviews for CHREBP Antibody (NB400-135) (5) 4.25

Average Rating: 4.2
(Based on 5 reviews)
We have 5 reviews tested in 3 species: Human, Mouse, Rat.
We have 5 reviews tested in 3 application: IHC-Fr, WB, ChIP.

Reviews using NB400-135:
Filter by Applications
IHC-Fr
(1)
WB
(3)
ChIP
(1)
All Applications
Filter by Species
Human
(1)
Mouse
(2)
Rat
(1)
All Species
Images Ratings Applications Species Date Details
Immunohistochemistry-Frozen CHREBP NB400-135
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reviewed by:
Anonymous
IHC-Fr Mouse 09/01/2017
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Summary

ApplicationImmunohistochemistry-Frozen
Sample Testedaortic tissues
SpeciesMouse
LotH1

Comments

CommentsThe frozen sections were permeabilized with 0.1% TritonX-100 and block with 10% goat serum. Sections were incubated with FITC-conjugated secondary antibodies.
 
reviewed by:
Yongjun Yu
WB Human 01/15/2013
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Summary

ApplicationWestern Blot
Sample Testedhuman fibroblast whole cell extract
SpeciesHuman
LotH-2

Blocking

Blocking Details5% nonfat milk in TBST, 30 min at RT

Primary Anitbody

Dilution Ratio1:200 in 2% BSA/TBST, overnight at 4C

Secondary Antibody

Secondary Descriptiongoat anti-rabbit IgG, HRP
Secondary Manufacturer Cat#Thermo Scientific, #31462
Secondary Concentration1:10,000

Details

Detection NotesLumi-light
 
reviewed by:
Deike Hesse
WB 12/13/2012
View

Summary

ApplicationWestern Blot
LotC2
Special ApplicationsUsed on mouse liver lysates. Detection in cytosol and nuclear fraction by Western blot.
Pub Med ID22927645
 
reviewed by:
Anonymous
ChIP Rat 11/28/2012
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Summary

ApplicationChromatin Immunoprecipitation
Sample TestedRat hepatocytes
SpeciesRat
CommentsWe found that this antibody worked very well for ChIP, however in our cells (primary rat hepatocytes) we could not detect ChREBP protein by Western blotting using this antibody. We are open to any suggestions that could help us detect ChREBP protein by Western blotting?

Primary Anitbody

Dilution RatioChREBP (6ug)

Details

Detection NotesRT-PCR
Fixation/Cross-Linking5M NaCl, 4hr incubation at 65 degrees Celsius
Immunoprecipitation DetailsProtein A agarose beads, 4hr incubation at 4 degrees Celcius
Crosslinking/Reversal Details1% formaldehyde
DNA Prurification DetailsPhenol chloroform

Comments

CommentsWe found that this antibody worked very well for ChIP, however in our cells (primary rat hepatocytes) we could not detect ChREBP protein by Western blotting using this antibody. We are open to any suggestions that could help us detect ChREBP protein by Western blotting?
Western Blot CHREBP NB400-135
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reviewed by:
Cathal McCarthy
WB Mouse 02/02/2011
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Summary

ApplicationWestern Blot
Sample TestedAorta whole tissue, Sample Amount: 20ug
SpeciesMouse
CommentsLoaded 20ug total protein, nitrocellulose membrane, blocked with milk, primary antibody dilution of 1:1000 and secondary dilution of 1;10000. Some non-specifics band at 75kDa also visible

Blocking

Blocking DetailsBlocking Buffer: 5% Milk in TBST, Blocking Time: 2 hours, Blocking Temp: Room temperature

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:1000 in milk, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Secondary Antibody

Secondary DescriptionSecondary Ab: Goat Anti-Mouse IgG HRP, Secondary Ab Dilution Ratio: 1:10000 in milk
Secondary Manufacturer Cat#sc-2005

Details

Detection NotesDetection Method: Supersignal Pierce, Exposure Time: 20 min, Specific Bands: 95kDa, Non-Specific Bands: 75kDa

Comments

CommentsLoaded 20ug total protein, nitrocellulose membrane, blocked with milk, primary antibody dilution of 1:1000 and secondary dilution of 1;10000. Some non-specifics band at 75kDa also visible

Product General Protocols

Video Protocols

WB Video Protocol
ChIP Video Protocol
ChIP Webinar
ICC/IF Video Protocol

FAQs for CHREBP Antibody (NB400-135). (Showing 1 - 1 of 1 FAQs).

  1. Pplease I want to block ChREBP Ab (NB400-135) using ChREBP peptide I need help to no the procedure and how much I need to add from the peptide to the ChREBP Ab
    • Here is a protocol for a blocking peptide control for a western blot: Materials and Reagents Blocking buffer (usually TBST plus either 5% non-fat dry milk or 3% BSA for Western blot, or PBS plus 1% BSA for IHC) - Antibody - Blocking (immunizing) peptide - Two tubes - Two identical samples (e.g. a Western blot with two identical lanes, cut in half; two slides containing the cells of interest; etc) Method 1. Determine the optimal concentration of antibody that consistently gives a positive result in your particular protocol. Using that concentration, determine how much antibody you will need for two experiments. a. For example, an antibody is being used successfully in Western blot at 0.5 ug/ml. You will need 2 ml of antibody solution to stain one strip of a Western blot. Thus, you would use 1 ug of antibody in 2 ml buffer for each strip. 2. Dilute the necessary amount of antibody in blocking buffer to the final volume needed for the two experiments. Divide this equally into two tubes. 3. In the first tube, labeled locked, add the blocking peptide to a final concentration of 1 ug/ml (2 ug total peptide in this example). In the second tube, labeled Control, add an equivalent amount of buffer. 4. Incubate both tubes, with agitation, at room temperature for 30 minutes, or overnight at 4 degrees C. 5. Perform the staining protocol on the two identical samples, using the blocked antibody for one and the control for the other. Be careful not to mix up the strips using the blocked and control antibodies! 6. Observe the staining. The staining that disappears when using the blocked antibody is specific to the antibody. (See note i) Notes i. If more than one band disappears in Western blot by peptide/antigen competition, those bands contain the antigenic determinants and could be fragments of the full antigen or a complex containing the antigen.

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB400-135AF405
Alexa Fluor 488 Labeled NB400-135AF488
Alexa Fluor 647 Labeled NB400-135AF647
Alexa Fluor 700 Labeled NB400-135AF700
Biotin Labeled NB400-135B
DyLight 405 Labeled NB400-135V
DyLight 488 Labeled NB400-135G
DyLight 550 Labeled NB400-135R
DyLight 650 Labeled NB400-135C
FITC Labeled NB400-135F
HRP Labeled NB400-135H

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Blogs on CHREBP.

ChREBP, a glucose sensitive transcription factor with role in glucose-lipids homeostasis and cancer
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...  Read full blog post.

ChREBP and Fatty Acid Biosynthesis Regulation
Carbohydrate response element binding protein (ChREBP) is a transcription factor involved in activating genes that encode enzymes of fatty acid biosynthesis in liver and adipose. It is believed to be a key controller of hepatic lipogenesis and has a p...  Read full blog post.

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Recent Reviews

4.2
5
1
4
4
3
0
2
0
1
0

Anonymous
09/01/2017
Application: IHC-Fr
Species: Mouse

 
Yongjun Yu
01/15/2013
Application: WB
Species: Human

 
Deike Hesse
12/13/2012
Application: WB
Species:

Bioinformatics

Gene Symbol MLXIPL