Revision 1

#79737Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-F, IF-IC, ChIP

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

50

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q8NHW3

Entrez-Gene Id:

389692

Product Information

Product Usage Information

For optimal ChIP results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
Immunofluorescence (Frozen) 1:1000
Immunofluorescence (Immunocytochemistry) 1:1000
Chromatin IP 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

MAFA (D6Z2N) recognizes endogenous levels of total MAFA protein. Based on sequence similarity, this antibody is not predicted to cross-react with MAFB or c-MAF.

Species Reactivity:

Human, Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala345 of human MAFA protein.

Background

MAFA belongs to the musculoaponeurotic fibrosarcoma (MAF) family of basic leucine-zipper transcription factors (1). In the mouse embryo, MAFA expression is first detected at E13.5, restricted to Nkx6.1-positive insulin-producing islet cells (2). Expression of the MAFA gene is sensitive to physiological glucose levels, and genomic targets regulated by MAFA include β-cell transcription factors (e.g., PDX1) and the insulin gene (2, 3). Ectopic expression of MAFA was shown to induce insulin production by pancreatic α-cells (2), while conditional overexpression of MAFA in vivo promoted transdifferentiation of α-cells into insulin-producing β-cells (4). Targeted deletion of the MAFA gene in mice likewise led to a loss of β-cell identity and function (5). Collectively, these data suggest that MAFA is critical for the development, maintenance, and physiological function of insulin-producing pancreatic β-cells, highlighting its potential utility as a target for translational and clinical research studies in diabetes (6).

  1. Hang, Y. and Stein, R. (2011) Trends Endocrinol Metab 22, 364-73.
  2. Matsuoka, T.A. et al. (2004) Proc Natl Acad Sci U S A 101, 2930-3.
  3. Vanhoose, A.M. et al. (2008) J Biol Chem 283, 22612-9.
  4. Matsuoka, T.A. et al. (2017) Diabetes 66, 1293-1300.
  5. Nishimura, W. et al. (2015) Diabetologia 58, 566-74.
  6. Lu, J. et al. (2017) Mol Med Rep 15, 4041-4048.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#79737

MAFA (D2Z6N) Rabbit mAb

Western Blotting Image 1: MAFA (D2Z6N) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using MAFA (D2Z6N) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). MAFA protein expression is detected exclusively in pancreatic beta cell lines (NIT-1 and Beta-TC-6), confirming target specificity of the antibody.
Western Blotting Image 2: MAFA (D2Z6N) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock-transfected (-) or transfected with Human MAFA (+), using MAFA (D2Z6N) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation Image 1: MAFA (D2Z6N) Rabbit mAb Expand Image
Immunoprecipitation of MAFA from NIT-1 cells. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is MAFA (D2Z6N) Rabbit mAb. Western blot analysis was performed using MAFA (D2Z6N) Rabbit mAb. Mouse Anti-Rabbit IgG (Conformation specific) (L27A9) mAb (HRP Conjugate) #5127 was used as the secondary antibody.
Immunofluorescence Image 1: MAFA (D2Z6N) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of adult mouse pancreas using MAFA (D2Z6N) Rabbit mAb (green pseudocolor). Insulin producing cells were labeled with Insulin (C27C9) Rabbit mAb (Alexa Fluor® 488 Conjugate) #9016 (red pseudocolor) after blocking free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).Note: Cross-linking with 4% formaldehyde should not exceed 4 hours; over-fixation diminishes the performance of this antibody.
Immunofluorescence Image 1: MAFA (D2Z6N) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of NIT-1 cells (left, positive) and NIH/3T3 cells (right, negative) using MAFA (D2Z6N) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red).
Chromatin Immunoprecipitation Image 1: MAFA (D2Z6N) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from Beta-TC-6 cells and either MAFA (D2Z6N) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using mouse insulin promoter primers, SimpleChIP® Mouse PDX1 Promoter Primers #60986, and SimpleChIP® Mouse GAPDH Intron 2 Primers #8986. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.