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Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb

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Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb

品牌： CST

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产品介绍

产品信息

荧光素标记

Unconjugated

抗原名称

SQSTM1p62

来源纯化

Monoclonal antibody is produced by immunizing animals with a synthetic phospho-peptide corresponding to residues surrounding Ser349 of human SQSTM1/62 protein.

宿主

Rabbit

商品描述

Product Usage Information

Application	Dilution
Western Blotting	1:1000
Immunohistochemistry (Paraffin)	1:1600 - 1:6400
Immunofluorescence (Immunocytochemistry)	1:200 - 1:800
Flow Cytometry (Fixed/Permeabilized)	1:400 - 1:1600

同种型

Rabbit IgG

分子量

内毒素水平

大鼠

研究领域

癌症,细胞生物学,纤维化,代谢,神经科学

应用

反应种属

Human,Mouse,

预测反应种属

Rat

目标/特异性

Specificity/Sensitivity

Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb recognizes endogenous levels of SQSTM1/p62 protein only when phosphorylated at Ser349. Staining of mitotic cells is observed by immunohistochemstry. The specificity of this staining is unknown.

Species Reactivity:

Human, Mouse

敏感性

Endogenous

背景

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.Phosphorylation of SQSTM1 at Ser349 (Ser351 in mouse) during oxidative stress increases its binding to KEAP1, thereby increasing NRF2 activity (13). 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. 13.Ichimura, Y. et al. (2013) Mol Cell 51, 618-31.

研究领域

癌症,细胞生物学,纤维化,代谢,神经科学

翻译后修饰

phosphate

制备和贮存

保存方式

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.

数据库链接

Entrez-Gene ID

8878

UniProt ID

Q13501

参考图片

Flow cytometric analysis of PC-3 cells, untreated (blue) or treated with sodium arsenite (15 µM, 18 hr; green), using Phospho-SQSTM1 (Ser349) (E7I4Z) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.

Western blot analysis of extracts from wild-type MEF and MEF SQSTM1/p62 KO cell lines, untreated (-) or treated with sodium arsenite (15 μM, 18 hr; +), using Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb (upper), SQSTM1/p62 (D1Q5S) Rabbit mAb #39749 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). MEF SQSTM1/p62 KO cells were kindly provided by Dr. Junying Yuan, Harvard Medical School, Boston MA.

Western blot analysis of extracts from PC-3 cells treated with sodium arsenite (15 μM, indicated times) using Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb (upper), SQSTM1/p62 (D5L7G) Mouse mAb #88588 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western blot analysis of extracts from PC-3 cells treated with sodium arsenite (15 μM, 18 hr) with lysates that were untreated (-) or treated with lambda-phosphatase and calf intestinal phosphatase (λPPase/CIP; +) using Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb (upper), SQSTM1/p62 (D5L7G) Mouse mAb #88588 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Immunohistochemical analysis of paraffin-embedded 293T cell pellet, untreated (left, negative) or treated with sodium arsenite (right, positive), using Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma, untreated (left) or lambda phosphatase treated (right), using Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma, untreated (left) or lambda phosphatase treated (right), using Phospho-SQSTM1/p62 (Ser349) (E7M1A) Rabbit mAb.

Confocal immunofluorescent analysis of PC-3 cells, untreated (left), treated with sodium arsenite (15 μM, 18 hr; center), or treated with sodium arsenite and post-processed with λ-phosphatase (right), using Phospho-SQSTM1 (Ser349) (E7M1A) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 650 Phalloidin #12956 (red). Blue = DAPI #4083 (fluorescent DNA dye).

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货号：

16177S

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