Revision 1

#59187Store at -20C

1 个试剂盒

(6 x 20 microliters)

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 Mol. Wt Isotype/Source
Beclin-1 (D40C5) Rabbit mAb 3495 20 µl 60 kDa Rabbit IgG
PI3 Kinase Class III (D9A5) Rabbit mAb 4263 20 µl 100 kDa Rabbit 
PIK3R4 Antibody 14580 20 µl 153 kDa Rabbit 
Atg14 (D1A1N) Rabbit mAb 96752 20 µl 65 kDa Rabbit IgG
UVRAG (D2Q1Z) Rabbit mAb 13115 20 µl 90 kDa Rabbit IgG
Rubicon (D9F7) Rabbit mAb 8465 20 µl 130 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Beclin-1 Complex Antibody Sampler Kit provides an economical means of detecting proteins that are part of the Beclin-1 complexes. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

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.

Background

A number of studies have identified distinct complexes involving Beclin-1 and PI3K Kinase Class III with specific roles in autophagy and vesicle trafficking (1,2). These complexes commonly contain Beclin-1, PI3KC3/VSP34, and PIK3R4/VPS15 and function to catalyze the phosphorylation of phosphatidylinositol at the D3 position, producing phosphatidylinositol-3-phosphate. Specificity of PI3KC3 activity is regulated by additional binding partners. Complex 1 contains Atg14 which is required for early stages of autophagosome nucleation (3,4). Complex 2 lacks Atg14, but instead contains UVRAG, and is important for autophagosome maturation and endocytic trafficking (4-6). A third complex, containing both UVRAG and Rubicon, negatively regulates canonical autophagy (7,8). Importantly, this complex containing Rubicon is critical for a related process of LC3-associated phagocytosis (LAP) in which extracellular pathogens binding to cell surface receptors are engulfed by a single membrane phagosome and degraded by the lysosome (9,10).

  1. Levine, B. and Kroemer, G. (2019) Cell 176, 11-42.
  2. Mei, Y. et al. (2016) Protein Sci 25, 1767-85.
  3. Sun, Q. et al. (2008) Proc Natl Acad Sci U S A 105, 19211-6.
  4. Itakura, E. et al. (2008) Mol Biol Cell 19, 5360-72.
  5. Liang, C. et al. (2006) Nat Cell Biol 8, 688-99.
  6. Liang, C. et al. (2008) Nat Cell Biol 10, 776-87.
  7. Matsunaga, K. et al. (2009) Nat Cell Biol 11, 385-96.
  8. Zhong, Y. et al. (2009) Nat Cell Biol 11, 468-76.
  9. Martinez, J. et al. (2015) Nat Cell Biol 17, 893-906.
  10. Heckmann, B.L. and Green, D.R. (2019) J Cell Sci 132(5). pii: jcs222984. doi: 10.1242/jcs.222984.

Background References

    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 1
    #59187

    Beclin-1 Complex Antibody Sampler Kit

    Beclin-1 Complex Antibody Sampler Kit: Image 1 Expand Image
    使用 UVRAG (D2Q1Z) Rabbit mAb(上图)或 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。HCT 116细胞被证实含有可导致 UVRAG 内源性表达水平显著降低的单等位基因移码突变 (11)。
    Beclin-1 Complex Antibody Sampler Kit: Image 2 Expand Image
    使用 PIK3R4 Antibody 对不同细胞系提取物进行蛋白质印迹分析。
    Beclin-1 Complex Antibody Sampler Kit: Image 3 Expand Image
    使用 Beclin-1 (D40C5) Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
    Beclin-1 Complex Antibody Sampler Kit: Image 4 Expand Image
    使用 PI3 Kinase Class III (D9A5) Rabbit mAb 对小鼠脑和 C6 细胞的提取物进行蛋白质印迹分析。
    Beclin-1 Complex Antibody Sampler Kit: Image 5 Expand Image
    一抗与靶标蛋白结合之后,与偶联 HRP 的二抗形成复合体。添加 LumiGLO®,在酶催化分解期间发光。
    Beclin-1 Complex Antibody Sampler Kit: Image 6 Expand Image
    使用 Rubicon (D9F7) Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
    Beclin-1 Complex Antibody Sampler Kit: Image 7 Expand Image
    使用 Atg14 (D1A1N) Rabbit mAb(上)和 β-Actin (D6A8) Rabbit mAb #84576(下)对 HCT 116 和 HCT 116/Atg14 shRNA 敲除型细胞提取物进行蛋白质印迹分析。HCT 116/Atg14 shRNA 细胞由 University of Minnesota, Minneapolis, MN Do-Hyung Kim 博士友情提供。
    Beclin-1 Complex Antibody Sampler Kit: Image 8 Expand Image
    使用 UVRAG (D2Q1Z) Rabbit mAb 对转染空载 (-) 或经构建表达的人 UVRAG (hUVRAG; +) 转染的 293T 细胞的提取物进行蛋白质印迹分析。
    Beclin-1 Complex Antibody Sampler Kit: Image 9 Expand Image
    使用 Beclin-1 (D40C5) XP® Rabbit mAb #3495(上)或 α-Tubulin (11H10) Rabbit mAb #2125(下)对经 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-)、SignalSilence® Beclin-1 siRNA I #6222 (+) 或 SignalSilence® Beclin-1 siRNA II (+) 转染的 Hela 细胞提取物进行蛋白质印迹分析。Beclin-1 (D40C5) XP® Rabbit mAb 可确认 Beclin-1 的表达沉默,而 α-Tubulin (11H10) Rabbit mAb 则用作观察上样量和 Beclin-1 siRNA 的特异性。
    Beclin-1 Complex Antibody Sampler Kit: Image 10 Expand Image
    使用 Rubicon (D9F7) Rabbit mAb 对转染空载 (-) 或经 人 Rubicon (hRubicon, +) 转染的 293T 细胞提取物进行蛋白质印迹分析。Rubicon 构建体由 University of California, Berkeley, CA 的 Qing Zhong 博士惠赠。
    Beclin-1 Complex Antibody Sampler Kit: Image 11 Expand Image
    使用 Atg14 (D1A1N) Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
    Beclin-1 Complex Antibody Sampler Kit: Image 12 Expand Image
    使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(泳道 2)或 UVRAG (D2Q1Z) Rabbit mAb(泳道 3)对 PANC-1 细胞提取物中的 UVRAG 进行免疫沉淀。泳道 1 是 10% 输入对照。使用 UVRAG (D2Q1Z) Rabbit mAb 进行蛋白质印迹分析。
    Beclin-1 Complex Antibody Sampler Kit: Image 13 Expand Image
    对 Neuro2A 细胞提取物 Atg14 进行免疫沉淀。泳道 1 为 10% 输入,泳道 2 用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 沉淀,泳道 3 为 Atg14 (D1A1N) Rabbit mAb。使用 Atg14 (D1A1N) Rabbit mAb 进行蛋白质印迹分析。