Overview

  • Product name
  • Description
    Rabbit polyclonal to MafA
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 300 to the C-terminus of Human MAFA.

    Read Abcam's proprietary immunogen policy (Peptide available as ab28523.)

  • Positive control
    • This antibody gave a positive signal in human brain tissue lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab26405 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 Use a concentration of 1 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 37 kDa).

Human MAFA has a predicted molecular weight of 37 kDa (Swiss-Prot data), however publications have shown the human protein to be detected at 47 kDa according to reference PMID 12011435, http://www.pnas.org/cgi/content/full/99/10/6737. Our data shows ab26405 to detect a band at 36 kDa in human brain, kidney and liver which is similar to that seen in other publications (PMID:14680841, PMID:18159220 (expression profile) ). The mouse protein has also been observed at 47 kDa according to reference PMID 12368292.

IHC-P Use a concentration of 0.8 - 4 µg/ml.
ICC/IF Use at an assay dependent concentration.

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

Images

  • The image shows human pancreas tissue in which some cells exhibit nuclear staining. The antigen retrieval method used was Tris EDTA pH9.
  • ab26405 staining MAFA in Human PANC-1 cell line transfected with synthetic mRNA by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.05% Triton X-100 and blocked with 5% serum for 30 minutes at 20°C. Samples were incubated with primary antibody (1/200) for 30 minutes at 37°C. An Alexa Fluor® 555-conjugated donkey anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.

    See Abreview

  • Anti-MafA antibody (ab26405) at 1 µg/ml + Brain (Human) Tissue Lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 37 kDa
    Observed band size: 36 kDa
    why is the actual band size different from the predicted?



    Human MAFA has a predicted molecular weight of 37 kDa (Swiss-Prot data).  Some publications suggest that the human protein should be detected at 47 kDa (PMID 12011435). Our data shows that ab26405 detects a band at 36 kDa in human brain, which is similar to that seen in other publications [PMID:14680841, PMID:18159220 (expression profile)]. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

References

This product has been referenced in:
  • Axelsson AS  et al. Sox5 regulates beta-cell phenotype and is reduced in type 2 diabetes. Nat Commun 8:15652 (2017). WB, IHC-P . Read more (PubMed: 28585545) »
  • Wang P  et al. A high-throughput chemical screen reveals that harmine-mediated inhibition of DYRK1A increases human pancreatic beta cell replication. Nat Med 21:383-8 (2015). WB, ICC/IF . Read more (PubMed: 25751815) »
See all 7 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (WT Black 6 mouse pancreas)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH 9.0
Permeabilization
No
Specification
WT Black 6 mouse pancreas
Fixative
10% Formalin

Dr. David Burk

Verified customer

Submitted Oct 05 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (pancreas)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA PH9
Permeabilization
No
Specification
pancreas
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jul 09 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Dorsal Root Ganglion)
Permeabilization
Yes - 0.2% Triton-X
Specification
Dorsal Root Ganglion
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Fixative
Paraformaldehyde

Irene Not_Provided

Verified customer

Submitted Dec 08 2017

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Sample
Mouse Tissue sections (Pancreas)
Specification
Pancreas
Permeabilization
Yes - Tween20 0.1%
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 08 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 for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C

Dr. Tomas Koblas

Verified customer

Submitted Mar 03 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 synthetic mRNA c)
Specification
PANC-1 cell line transfected with synthetic mRNA c
Permeabilization
Yes - 0.05 % Triton X100
Fixative
Formaldehyde

Dr. Tomas Koblas

Verified customer

Submitted Feb 24 2014

Answer

Thank you for contacting us. The IHC image on the datasheet for ab26405 was provided by a collaborator and we unfortunately do not have their exact protocol. However, it is safe to assume that the protocol is standard and that the unmasking procedure was similar to the procedure described in the PDF file I have attached, in section D: Antigen Retrieval. This general protocol can also be found on our website at this link: https://www.abcam.com/ps/pdf/protocols/ihc_p.pdf Section D contains a recipe for Tris-EDTA pH 9.0 and recommendations for heating. Briefly, if using a microwave or steamer or water bath, you will want to maintain the slides in the buffer at 95-100C for 10 - 20 minutes, followed by cooling for another 20 minutes. The optimal amount of time for your tissue sections will need to be determined but in general, 20 minutes at 95-100C is usually sufficient. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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