Revision 3

#2895Store 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-IC, FC-FP, ChIP

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

27

Source/Isotype:

Mouse IgG2a

UniProt ID:

#P35638

Entrez-Gene Id:

1649

Product Information

Product Usage Information

For optimal ChIP results, use 2.5 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 1:50
Immunofluorescence (Immunocytochemistry) 1:400 - 1:1600
Flow Cytometry (Fixed/Permeabilized) 1:250 - 1:1000
Chromatin IP 1:200

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.

For a carrier free (BSA and azide free) version of this product see product #78063.

Specificity / Sensitivity

CHOP (L63F7) Mouse mAb detects endogenous levels of total CHOP protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the sequence of human CHOP.

Background

CHOP was identified as a C/EBP-homologous protein that inhibits C/EBP and LAP in a dominant-negative manner (1). CHOP expression is induced by certain cellular stresses including starvation and the induced CHOP suppresses cell cycle progression from G1 to S phase (2). Later it was shown that, during ER stress, the level of CHOP expression is elevated and CHOP functions to mediate programmed cell death (3). Studies also found that CHOP mediates the activation of GADD34 and Ero1-Lα expression during ER stress. GADD34 in turn dephosphorylates phospho-Ser51 of eIF2α thereby stimulating protein synthesis. Ero1-Lα promotes oxidative stress inside the endoplasmic reticulum (ER) (4). The role of CHOP in the programmed cell death of ER-stressed cells is correlated with its role promoting protein synthesis and oxidative stress inside the ER (4).

  1. Ron, D. and Habener, J.F. (1992) Genes Dev 6, 439-53.
  2. Barone, M.V. et al. (1994) Genes Dev 8, 453-64.
  3. Zinszner, H. et al. (1998) Genes Dev 12, 982-95.
  4. Marciniak, S.J. et al. (2004) Genes Dev 18, 3066-77.

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-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) 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.
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 3
#2895

CHOP (L63F7) Mouse mAb

Western Blotting Image 1: CHOP (L63F7) Mouse mAb Expand Image
使用 CHOP (L63F7) Mouse mAb 对未处理的或经 Thapsigargin (300 nM,2 小时)或 Tunicamycin(24 μg/ml,2 小时)处理的 C6 和 A-204 细胞提取物进行蛋白质印迹法分析。
No image available
Immunofluorescence Image 1: CHOP (L63F7) Mouse mAb Expand Image
使用 CHOP (L63F7) Mouse mAb(绿色)对未处理的(左)或经 Tunicamycin 处理(右)的 A-204 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 DY-554 Phalloidin 进行标记(红色)。
Flow Cytometry Image 1: CHOP (L63F7) Mouse mAb Expand Image
使用 CHOP (L63F7) 小鼠单克隆抗体(实线)或浓度匹配的小鼠 (E5Y6Q) 单克隆抗体 IgG2a 同型对照 #61656(虚线)对未经(蓝色)或已经 tunicamycin(2 µg/mL,8 小时;绿色)处理的 HeLa 细胞进行流式细胞分析。使用 Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 作为二抗。
Chromatin Immunoprecipitation Image 1: CHOP (L63F7) Mouse mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 对经衣霉素 (2 ug/ml) 过夜处理的 MEF 野生型细胞的交联染色质和 CHOP (L63F7) Mouse mAb #2895 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀分析。使用 SimpleChIP® Mouse ATF-3 Intron 1 Primers #13059、Mouse CHOP Promoter Primers 和 SimpleChIP® Mouse RPL30 Intron 2 Primers #7015,通过实时 PCR 对富集的 DNA 进行定量。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。