Overview

  • Product name
    Anti-MafA antibody - ChIP Grade
    See all MafA primary antibodies
  • Description
    Rabbit polyclonal to MafA - ChIP Grade
  • Specificity
    Antibody has not been used successfully for the human ortholog.
  • Tested applications
    Suitable for: WB, ChIP, EMSA, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse
    Predicted to work with: Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Mouse MafA (C terminal).
    Database link: 389692

Properties

Applications

Our Abpromise guarantee covers the use of ab17976 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/10000.
ChIP Use 5-15µg for 108 cells.
EMSA Use a concentration of 0.05 - 0.25 mg/ml.
IHC-P Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-Fr 1/1000 - 1/3000.

Target

  • Function
    Acts as a transcriptional factor. Specifically binds the insulin enhancer element RIPE3b and activates insulin gene expression. Cooperates synergistically with NEUROD1 and PDX1. Phosphorylation by GSK3 increases its transcriptional activity and is required for its oncogenic activity. Involved either as an oncogene or as a tumor suppressor, depending on the cell context.
  • Sequence similarities
    Belongs to the bZIP family. Maf subfamily.
    Contains 1 bZIP domain.
  • Post-translational
    modifications
    Ubiquitinated, leading to its degradation by the proteasome.
    Ser-14 and Ser-65 appear to be the major phosphorylation sites. Phosphorylated by MAPK13 on serine and threonine residues (Probable). Phosphorylation by GSK3 requires prior phosphorylation of Ser-65 by another kinase. Phosphorylation proceeds then from Ser-61 to Thr-57, Thr-53 and Ser-49. GSK3-mediated phosphorylation increases its transcriptional activity through the recruitment of the coactivator PCAF, is required for its transforming activity and leads to its degradation through an ubiquitin/proteasome-dependent pathway.
  • Cellular localization
    Nucleus. Detected in nuclei of pancreas islet beta cells.
  • Information by UniProt
  • Database links
  • Alternative names
    • HMafA antibody
    • Mafa antibody
    • MAFA_HUMAN antibody
    • Pancreatic beta cell specific transcriptional activator antibody
    • Pancreatic beta-cell-specific transcriptional activator antibody
    • RIPE3b1 antibody
    • Transcription factor MafA antibody
    • Transcription factor RIPE3b1 antibody
    • v maf musculoaponeurotic fibrosarcoma oncogene homolog A (avian) antibody
    • v maf musculoaponeurotic fibrosarcoma oncogene homolog A antibody
    • V-maf musculoaponeurotic fibrosarcoma oncogene homolog A antibody
    see all

Anti-MafA antibody - ChIP Grade images

  • Binding of MafA to the enhancer region of the endogenous insulin gene. Samples: Immunoprecipitated cross-linked DNA from betaTC-3 cells was analyzed by PCR. As controls, reactions were run with no DNA, with input chromatin, with DNA obtained after precipitation with rabbit IgG or without antibody (lanes 1 through 5, respectively). Antibody: Affinity purified rabbit anti-MafA antibody. Test

  • Electrophoretic Mobility Shift of MafA, MafB and c-Maf. Samples: Nuclear extract (6 ug) from HeLa cells transfected with MafA, MafB or c-Maf expression constructs. Antibodies: Affinity purified anti-MafA antibody BL1069, anti-MafB Antibody or anti-c-Maf antibody.

  • ab17976 staining MAFA in Mouse pancreas tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 20% serum for 30 minutes at 22°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/2000 in blocking buffer) for 16 hours at 22°C. A Biotin-conjugated Donkey anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.

    See Abreview

  • Detection of MafA, MafB and cMaf by Western Blot. Samples: Nuclear extract (6 ug) from HeLa cells transfected with MafA, MafB or cMaf expression constructs. Antibodies: Affinity purified anti-MafA antibody, anti-MafB Antibody or anti-c-Maf antibody. Each antibody was used at 0.5 ug/ml. Detection: Chemiluminescence with a 5 second exposure.

References for Anti-MafA antibody - ChIP Grade (ab17976)

This product has been referenced in:
  • Roth Flach RJ  et al. Protein Kinase Mitogen-activated Protein Kinase Kinase Kinase Kinase 4 (MAP4K4) Promotes Obesity-induced Hyperinsulinemia. J Biol Chem 291:16221-30 (2016). IHC-P ; Mouse . Read more (PubMed: 27226575) »
  • Sakano D  et al. VMAT2 identified as a regulator of late-stage ß-cell differentiation. Nat Chem Biol 10:141-8 (2014). Read more (PubMed: 24316738) »

See all 4 Publications for this product

Product Wall

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Pancreas, Islets of Langerhans, beta-cells)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Bond Epitope Retrieval Solution 2
Permeabilization
No
Specification
Pancreas, Islets of Langerhans, beta-cells
Blocking step
hydrogen peroxide as blocking agent for 5 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Formaldehyde
Username

Reggie Williams

Verified customer

Submitted Aug 05 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3%
Sample
Rat Cell (Insulinoma cell line INS-1)
Specification
Insulinoma cell line INS-1
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Fixative
Paraformaldehyde
Username

Dr. Javier Triñanes

Verified customer

Submitted Jan 19 2015

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (12)
Sample
Human Cell lysate - whole cell (Human pancreatic Islets and human PANC-1 cell line)
Specification
Human pancreatic Islets and human PANC-1 cell line
Blocking step
ROCHE Western Blocking Reagent as blocking agent as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Dr. Tomas Koblas

Verified customer

Submitted Feb 26 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Sample
Human Cell (PANC-1 cell line transfected with mRNA coding for)
Specification
PANC-1 cell line transfected with mRNA coding for
Permeabilization
Yes - 0.05 % Triton X100
Fixative
Formaldehyde
Username

Dr. Tomas Koblas

Verified customer

Submitted Feb 26 2014

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (pancreas)
Specification
pancreas
Fixative
Formaldehyde
Antigen retrieval step
None
Permeabilization
No
Blocking step
(agent) for 10 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Ms. sarah shen

Verified customer

Submitted May 15 2013

Thank you for your reply and the nice feedback.
I have checked our stock, and it seems we only have the same lot number available you had previously. I can assure you that you have our Abpromise on every replacement as well if this one would fail ...

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Thank you for taking time to answer my questions.
Having reviewed the additional protocol details, I believe this product should have given satisfactory results. It appears that you may have received two faulty vials, which is regrettable as this ...

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Thank you for contacting us.
I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.
I am attaching ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (mouse pancreas)
Specification
mouse pancreas
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 20% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Jul 30 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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