Revision 1

#69375Store 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, IF-IC, ChIP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

70

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q12857

Entrez-Gene Id:

4774

Product Information

Product Usage Information

For optimal ChIP results, use 10 μL of antibody and 10 μg of chromatin (approximately 4 × 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
Application Dilution
Western Blotting 1:1000
Immunofluorescence (Immunocytochemistry) 1:50 - 1:200
Chromatin IP 1:50

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

NFI-A (E6Z1R) Rabbit mAb recognizes endogenous levels of total NFI-A protein. Based on sequence homology, this antibody is not predicted to cross-react with other NFI family members.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro430 of human NFI-A protein.

Background

NFI-A belongs to the nuclear factor I (NFI) family of site-specific transcription factors that regulate viral DNA replication and expression of various genes (1,2). The NFI family is composed of four members in vertebrates: NFI-A, NFI-B, NFI-C, and NFI-X, all of which are critical in the development of multiple organ systems in mice and humans (3). NFI-A activates cell-type-specific enhancers and facilitates the binding of PPARγ, the master transcriptional regulator of adipogenesis, to control the brown fat gene program (4). Additionally, NFI-A represses the expression of Myod1, the master transcriptional regulator of myogenesis, thus actively suppressing the myogenic gene program (5).

  1. Gronostajski, R.M. (2000) Gene 249, 31-45.
  2. Liang, B. et al. (2012) J Virol 86, 3264-75.
  3. Steele-Perkins, G. et al. (2003) Mol Cell Biol 23, 1075-84.
  4. Hiraike, Y. et al. (2017) Nat Cell Biol 19, 1081-1092.
  5. Hiraike, Y. et al. (2020) PLoS Genet 16, e1009044.

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 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.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
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
#69375

NFI-A (E6Z1R) Rabbit mAb

Western Blotting Image 1: NFI-A (E6Z1R) Rabbit mAb Expand Image
使用 NFI-A (E6Z1R) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对各种细胞系的提取物进行蛋白质印迹法分析。NFI-A 蛋白在 JHH-1 细胞中的低表达与预测的表达模式一致。
Immunofluorescence Image 1: NFI-A (E6Z1R) Rabbit mAb Expand Image
使用 NFI-A (E6Z1R) Rabbit mAb(绿色)、DyLight 650 Phalloidin #12956(红色)和 DAPI #4083(蓝色)对 JHH-2 细胞(左图,阳性)和 JHH-1 细胞(右图,阴性)进行共聚焦免疫荧光分析。
Chromatin Immunoprecipitation Image 1: NFI-A (E6Z1R) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对未经处理(左小图)或血清饥饿 72 小时(右小图)的 GM16633 细胞的交联染色质和 NFI-A (E6Z1R) Rabbit mAb 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀法分析。使用人 p21 启动子引物和人 p21 上游引物,通过实时 PCR 对富集的 DNA 进行定量分析。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。