Revision 5

#23214Store 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, IHC-P, IF-IC

REACTIVITY:

M R

SENSITIVITY:

Endogenous

MW (kDa):

62

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q64337

Entrez-Gene Id:

18412

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
Immunohistochemistry (Paraffin) 1:125 - 1:500
Immunofluorescence (Immunocytochemistry) 1:400 - 1:1600

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

SQSTM1/p62 (D6M5X) Rabbit mAb recognizes endogenous levels of total rodent SQSTM1/p62 protein.

Species Reactivity:

Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly300 of mouse SQSTM1/p62 protein.

Background

Sequestosome 1 (SQSTM1, p62) is a ubiquitin binding protein involved in cell signaling, oxidative stress, and autophagy (1-4). It was first identified as a protein that binds to the SH2 domain of p56Lck (5) and independently found to interact with PKCζ (6,7). SQSTM1 was subsequently found to interact with ubiquitin, providing a scaffold for several signaling proteins and triggering degradation of proteins through the proteasome or lysosome (8). Interaction between SQSTM1 and TRAF6 leads to the K63-linked polyubiquitination of TRAF6 and subsequent activation of the NF-κB pathway (9). Protein aggregates formed by SQSTM1 can be degraded by the autophagosome (4,10,11). SQSTM1 binds autophagosomal membrane protein LC3/Atg8, bringing SQSTM1-containing protein aggregates to the autophagosome (12). Lysosomal degradation of autophagosomes leads to a decrease in SQSTM1 levels during autophagy; conversely, autophagy inhibitors stabilize SQSTM1 levels. Studies have demonstrated a link between SQSTM1 and oxidative stress. SQSTM1 interacts with KEAP1, which is a cytoplasmic inhibitor of NRF2, a key transcription factor involved in cellular responses to oxidative stress (3). Thus, accumulation of SQSTM1 can lead to an increase in NRF2 activity.

  1. Kirkin, V. et al. (2009) Mol Cell 34, 259-69.
  2. Seibenhener, M.L. et al. (2007) FEBS Lett 581, 175-9.
  3. Komatsu, M. et al. (2010) Nat Cell Biol 12, 213-23.
  4. Bjørkøy, G. et al. (2006) Autophagy 2, 138-9.
  5. Joung, I. et al. (1996) Proc Natl Acad Sci USA 93, 5991-5.
  6. Sanchez, P. et al. (1998) Mol Cell Biol 18, 3069-80.
  7. Puls, A. et al. (1997) Proc Natl Acad Sci USA 94, 6191-6.
  8. Vadlamudi, R.K. et al. (1996) J Biol Chem 271, 20235-7.
  9. Wooten, M.W. et al. (2005) J Biol Chem 280, 35625-9.
  10. Bjørkøy, G. et al. (2005) J Cell Biol 171, 603-14.
  11. Komatsu, M. et al. (2007) Cell 131, 1149-63.
  12. Pankiv, S. et al. (2007) J Biol Chem 282, 24131-45.

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 IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry)

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.
SignalStain is a registered trademark of Cell Signaling Technology, Inc.
XP 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 5
#23214

SQSTM1/p62 (D6M5X) Rabbit mAb

Western Blotting Image 1: SQSTM1/p62 (D6M5X) Rabbit mAb Expand Image
使用 SQSTM1/p62 (D6M5X) Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对来自野生型小鼠或 SQSTM1/p62 敲除型小鼠的 MEF 细胞的提取物进行蛋白质印迹分析。MEF SQSTM1/p62 KO 细胞由马萨诸塞州波士顿哈佛大学医学院的 Junying Yuan 博士提供。
Western Blotting Image 2: SQSTM1/p62 (D6M5X) Rabbit mAb Expand Image
使用 SQSTM1/p62 (D6M5X) Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对未经处理 (-) 或经Chloroquine #14774(50 μM,过夜)处理的 C2C12 细胞的提取物进行蛋白质印迹分析。
Western Blotting Image 3: SQSTM1/p62 (D6M5X) Rabbit mAb Expand Image
使用 SQSTM1/p62 (D6M5X) Rabbit mAb 对不同细胞系的提取物进行蛋白质印迹分析。
Western Blotting Image 4: SQSTM1/p62 (D6M5X) Rabbit mAb Expand Image
使用 SQSTM1/p62 (D6M5X) Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对未经处理 (-) 或经 Earles 碱性盐溶液(EBSS;4 小时;+)处理的 MEF 细胞的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: SQSTM1/p62 (D6M5X) Rabbit mAb Expand Image
对 L-929 细胞提取物的 QSTM1 进行免疫沉淀。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 SQSTM1/p62 (D6M5X) Rabbit mAb。使用 SQSTM1/p62 (D6M5X) Rabbit mAb 进行蛋白质印迹法分析。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 用作二抗。
Immunohistochemistry Image 1: SQSTM1/p62 (D6M5X) Rabbit mAb Expand Image
使用 SQSTM1/p62 (D6M5X) Rabbit mAb 对石蜡包埋的 MEF 野生型细胞沉淀物(左图,阳性)或 MEF SQSTM1/p62 KO 细胞沉淀物(右图,阴性)进行免疫组织化学分析。MEF SQSTM1/p62 KO 细胞由马萨诸塞州波士顿哈佛大学医学院的 Junying Yuan 博士提供。
Immunohistochemistry Image 2: SQSTM1/p62 (D6M5X) Rabbit mAb Expand Image
使用 SQSTM1/p62 (D6M5X) Rabbit mAb 对石蜡包埋的小鼠前胃组织进行免疫组织化学分析。
Immunohistochemistry Image 3: SQSTM1/p62 (D6M5X) Rabbit mAb Expand Image
使用 SQSTM1/p62 (D6M5X) Rabbit mAb 对石蜡包埋的小鼠肾脏组织进行免疫组织化学分析。
Immunohistochemistry Image 4: SQSTM1/p62 (D6M5X) Rabbit mAb Expand Image
使用 SQSTM1/p62 (D6M5X) Rabbit mAb 对石蜡包埋的小鼠脾脏组织进行免疫组织化学分析。
Immunohistochemistry Image 5: SQSTM1/p62 (D6M5X) Rabbit mAb Expand Image
使用 SQSTM1/p62 (D6M5X) Rabbit mAb 对石蜡包埋的大鼠脾脏组织进行免疫组织化学分析。
Immunohistochemistry Image 6: SQSTM1/p62 (D6M5X) Rabbit mAb Expand Image
使用 SQSTM1/p62 (D6M5X) Rabbit mAb 对石蜡包埋的小鼠小肠组织进行免疫组织化学分析。
Immunofluorescence Image 1: SQSTM1/p62 (D6M5X) Rabbit mAb Expand Image
使用 SQSTM1 (D6M5X) Rabbit mAb(绿色)对未经(左图)或已经 Chloroquine #14774(50 μM,18 小时;中间)处理的野生型 MEF 和经 氯喹 处理(右图)的 SQSTM1/p62 敲除型 MEF 进行共聚焦免疫荧光分析。肌动蛋白丝用 β-Actin (8H10D10) Mouse mAb #3700(红色)标记。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。MEF SQSTM1/p62 KO 细胞由马萨诸塞州波士顿哈佛大学医学院的 Junying Yuan 博士提供。