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.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 1
#79737

MAFA (D2Z6N) Rabbit mAb

Western Blotting Image 1: MAFA (D2Z6N) Rabbit mAb Expand Image
使用 MAFA (D2Z6N) Rabbit mAb(上图)和 β-Actin (D6A8) Rabbit mAb #8457(下图),对不同细胞系提取物进行蛋白质印迹分析。MAFA 蛋白表达只在胰腺 β 细胞系(NIT-1 和 β-TC-6)中检测到,这确认了抗体的靶标特异性。
Western Blotting Image 2: MAFA (D2Z6N) Rabbit mAb Expand Image
使用 MAFA (D2Z6N) Rabbit mAb(上图)和 β-Actin (D6A8) Rabbit mAb #8457(下图)对转染空载 (-) 或使用人 MAFA (+) 转染的 293T 细胞提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: MAFA (D2Z6N) Rabbit mAb Expand Image
对 NIT-1 细胞的 MAFA 进行免疫沉淀。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 MAFA (D2Z6N) Rabbit mAb。使用 MAFA (D2Z6N) Rabbit mAb 进行蛋白质印迹分析。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 用作二抗。
Immunofluorescence Image 1: MAFA (D2Z6N) Rabbit mAb Expand Image
使用 MAFA (D2Z6N) Rabbit mAb(绿色假色)对成体小鼠胰腺细胞进行共聚焦免疫荧光分析。使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 封闭了游离二抗结合位点之后,用 Insulin (C27C9) Rabbit mAb (Alexa Fluor® 488 Conjugate) #9016(红色假色)标记胰岛素生成细胞。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。注:与 4% 甲醛交联不应超过 4 小时;过度固定会减弱该抗体的性能。
Immunofluorescence Image 1: MAFA (D2Z6N) Rabbit mAb Expand Image
使用 MAFA (D2Z6N) Rabbit mAb (Mouse Specific)(绿色)对 NIT-1 细胞(左图,阳性)和 NIH/3T3 细胞(右图,阴性)进行共聚焦免疫荧光分析。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)标记。
Chromatin Immunoprecipitation Image 1: MAFA (D2Z6N) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 β-TC-6 细胞中提取的交联染色质,在加入 MAFA (D2Z6N) Rabbit mAb 或 Normal Rabbit IgG #2729 后,进行染色质免疫沉淀。使用小鼠胰岛素启动子引物、 SimpleChIP® Mouse PDX1 Promoter Primers #60986 和 SimpleChIP® Mouse GAPDH Intron 2 Primers #8986.,通过实时 PCR 对富集 DNA 进行量化。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量(等于 1)的信号。