Revision 1

#7185Store at +4C

 

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Species Cross Reactivity

H M

UniProt ID:

#P15336

Entrez-Gene Id:

#1386

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.
Product Includes Product # Quantity Color Storage Temp
ATF2 Mouse Detection mAb 7186 1 ea Green (Lyophilized) +4C
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) 13304 1 ea Red (Lyophilized) +4C
Detection Antibody Diluent 13339 11 ml Green +4C
HRP Diluent 13515 11 ml Red +4C
TMB Substrate 7004 11 ml Colorless +4C
STOP Solution 7002 11 ml Colorless +4C
Sealing Tape 54503 2 ea +4C
ELISA Wash Buffer (20X) 9801 25 ml Colorless +4C
ELISA Sample Diluent 11083 25 ml Blue +4C
Cell Lysis Buffer (10X) 9803 15 ml Yellowish -20C

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

CST's PathScan® Phospho-ATF-2 (Thr71) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-ATF-2 (Thr71) protein. A Phospho-ATF-2 (Thr 71) Antibody has been coated onto the microwells. After incubation with cell lysates, phospho-ATF-2 protein is captured by the coated antibody. Following extensive washing, a total ATF-2 Mouse mAb is added to detect the captured phospho-ATF-2 protein. HRP-linked Anti-mouse Antibody #7076 is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of phospho-ATF-2 (Thr71).
Antibodies in kit are custom formulations specific to kit.

Specificity/Sensitivity

CST's PathScan® Phospho-ATF-2 (Thr71) Sandwich ELISA Kit detects endogenous levels of Phospho-ATF-2 protein. Using this Sandwich ELISA Kit #7185, a significant induction of phospho-ATF-2 in NIH/3T3 cells treated with anisomycin can be detected. However, the level of total ATF-2 (phospho- and nonphospho-), detected by PathScan® Total ATF-2 Sandwich ELISA Kit #7195, remains unchanged (Figure 1). This kit can also be used to detect phosphorylated ATF-2 protein in human HeLa and 293 cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

The transcription factor ATF-2 (also called CRE-BP1) binds to both AP-1 and CRE DNA response elements and is a member of the ATF/CREB family of leucine zipper proteins (1). ATF-2 interacts with a variety of viral oncoproteins and cellular tumor suppressors and is a target of the SAPK/JNK and p38 MAP kinase signaling pathways (2-4). Various forms of cellular stress, including genotoxic agents, inflammatory cytokines, and UV irradiation, stimulate the transcriptional activity of ATF-2. Cellular stress activates ATF-2 by phosphorylation of Thr69 and Thr71 (2-4). Both SAPK and p38 MAPK have been shown to phosphorylate ATF-2 at these sites in vitro and in cells transfected with ATF-2. Mutations of these sites result in the loss of stress-induced transcription by ATF-2 (2-4). In addition, mutations at these sites reduce the ability of E1A and Rb to stimulate gene expression via ATF-2 (2).

  1. Abdel-Hafiz, H.A. et al. (1992) Mol Endocrinol 6, 2079-89.
  2. Gupta, S. et al. (1995) Science 267, 389-93.
  3. van Dam, H. et al. (1995) EMBO J 14, 1798-811.
  4. Livingstone, C. et al. (1995) EMBO J 14, 1785-97.
  5. Emre, Y. et al. (2007) Biochem J 402, 271-8.
  6. Ipaktchi, K. et al. (2006) J Immunol 177, 8065-71.

Background References

    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.
    PathScan 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

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    Revision 1
    #7185

    PathScan® Phospho-ATF-2 (Thr71) Sandwich ELISA kit

    PathScan® Phospho-ATF-2 (Thr71) Sandwich ELISA kit: Image 1 Expand Image
    图 2:未经和已经过茴香霉素处理的 NIH/3T3 细胞的裂解物蛋白浓度与使用试剂盒测得的光密度读数之间存在关系。NIH/3T3 细胞(80% 融合度)用 茴香霉素 处理,并在 37ºC 下孵育 30 分钟后进行裂解。
    PathScan® Phospho-ATF-2 (Thr71) Sandwich ELISA kit: Image 2 Expand Image
    图 1:NIH/3T3 细胞用茴香霉素处理会刺激 ATF-2 在 Thr71 位点的磷酸化,使用 PathScan® Phospho-ATF-2 (Thr71) Sandwich ELISA kit #7185 可进行检测,但不影响 PathScan® Total ATF-2 ELISA kit #7195 检测的 ATF-2 总蛋白的水平。OD 450 读数如图的上部所示,使用 ATF-2 Rabbit mAb #9226(左小图)或 phospho-ATF-2 (Thr71) Antibody #9221(右小图)检测到的相应蛋白质印迹如图的底部所示。