Revision 1

#11874Store at +4C

 

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Species Cross Reactivity

H

UniProt ID:

#P10415

Entrez-Gene Id:

#596

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
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

The PathScan® Phospho-Bcl-2 (Ser70) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Bcl-2 when phosphorylated at Ser70. A Phospho-Bcl-2 (Ser70) Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, phosphorylated Bcl-2 protein is captured by the coated antibody. Following extensive washing, a Bcl-2 Mouse Detection mAb is added to detect the captured phospho-Bcl-2 (Ser70) protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for the developed color is proportional to the quantity of Bcl-2 phosphorylated at Ser70.
Antibodies in kit are custom formulations specific to kit.

Specificity/Sensitivity

The PathScan® Phospho-Bcl-2 (Ser70) Sandwich ELISA Kit recognizes endogenous levels of Bcl-2 protein when phosphorylated at Ser70 as shown in Figure 1. Kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

Bcl-2 exerts a survival function in response to a wide range of apoptotic stimuli through inhibition of mitochondrial cytochrome c release (1). It has been implicated in modulating mitochondrial calcium homeostasis and proton flux (2). Several phosphorylation sites have been identified within Bcl-2, including Thr56, Ser70, Thr74, and Ser87 (3). It has been suggested that these phosphorylation sites may be targets of the ASK1/MKK7/JNK1 pathway and that phosphorylation of Bcl-2 may be a marker for mitotic events (4,5). Mutation of Bcl-2 at Thr56 or Ser87 inhibits its anti-apoptotic activity during glucocorticoid-induced apoptosis of T lymphocytes (6). Interleukin-3 and JNK-induced Bcl-2 phosphorylation at Ser70 may be required for its enhanced anti-apoptotic functions (7).

  1. Murphy, K.M. et al. (2000) Cell Death Differ 7, 102-11.
  2. Zhu, L. et al. (1999) J Biol Chem 274, 33267-73.
  3. Maundrell, K. et al. (1997) J Biol Chem 272, 25238-42.
  4. Yamamoto, K. et al. (1999) Mol Cell Biol 19, 8469-78.
  5. Ling, Y.H. et al. (1998) J Biol Chem 273, 18984-91.
  6. Huang, S.T. and Cidlowski, J.A. (2002) FASEB J 16, 825-32.
  7. Deng, X. et al. (2001) J Biol Chem 276, 23681-8.

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

    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
    #11874

    PathScan® Phospho-Bcl-2 (Ser70) Sandwich ELISA Kit

    PathScan® Phospho-Bcl-2 (Ser70) Sandwich ELISA Kit: Image 1 Expand Image
    图 2. 未经处理或经 λ 磷酸酶或经 Paclitaxel #9807(1 mM,20 小时;37ºC)处理的 Jurkat 细胞的裂解物蛋白浓度与在 450 nm 处的吸光度之间的关系如图所示。
    PathScan® Phospho-Bcl-2 (Ser70) Sandwich ELISA Kit: Image 2 Expand Image
    图 1. Jurkat 细胞用 Paclitaxel 处理会刺激Bcl-2在Ser70位点的磷酸化,使用 PathScan® Phospho-Bcl-2 (Ser70) Sandwich ELISA Kit 可进行检测,但不影响 PathScan® Total Bcl-2 Sandwich ELISA Kit #12030 检测的总 Bcl-2 的水平。Jurkat 细胞未经处理或用 λ 磷酸酶或 Paclitaxel #9807(1 mM,20 小时;37ºC)处理。在 450 nm 处的吸光度读数如顶部图所示,使用 Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb #2827(左小图)或 Bcl-2 (D55G8) Rabbit mAb (Human Specific) #4223(右小图)检测到的相应蛋白质印迹如底部图所示。