Cell Signaling Technology

Product Pathways - Protein Stability

DUB Antibody Sampler Kit #8353

REACTIVITY

No. Size Price
8353T 1 Kit ( 9 x 20 µl ) ¥7,245.00 现货查询 购买询价
Kit Includes Quantity Applications Reactivity Homology† MW (kDa) Isotype
UCHL1 (D3T2E) XP® Rabbit mAb #13179 20 µl W,IHC-P,IF-IC,F, H,M,R,Mk, Hm,B,Dg,Pg,Hr, 27 Rabbit IgG
Phospho-CYLD (Ser418) Antibody #4500 20 µl W, H, 108 Rabbit
HAUSP (D17C6) XP® Rabbit mAb #4833 20 µl W,IP,IF-IC, H,M,R,Mk, 135, 140 Rabbit IgG
STAMBP Antibody #5245 20 µl W,IP, H,Mk, 50 Rabbit
A20/TNFAIP3 (D13H3) Rabbit mAb #5630 20 µl W,IP, H,M,R,Mk, 82 Rabbit IgG
USP9X Antibody #5751 20 µl W, H,M,R,Mk,Dg, 270 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl W, Goat
UCHL3 (D25E6) Rabbit mAb #8141 20 µl W, H,M,R, B,Dg,Hr, 27 Rabbit IgG
CYLD (D1A10) Rabbit mAb #8462 20 µl W,IP, H,M,R,Mk, Hm,B,Dg,GP,Hr, 108 Rabbit IgG
USP10 (D7A5) Rabbit mAb #8501 20 µl W,IP,IF-IC, H,M,R,Mk, 110 Rabbit IgG

Specificity / Sensitivity

Phospho-CYLD (Ser418)抗体识别内源性的Ser418位磷酸化CYLD蛋白。这个试剂盒的其他抗体用于目的蛋白的总蛋白水平检测。USP9X抗体可能会与USP9Y有交叉反应。

Source / Purification

Phospho-CYLD (Ser418)通过合成对应于人CYLD蛋白Ser418位点周围序列的磷酸化肽段免疫动物获得。试剂盒中其他多克隆抗体是分别通过合成对应于人CYLD蛋白His334位点、人STAMBP的Gly194位点、人UCHL1的Asp110位点、人UCHL3的Leu220位点、人USP10的Leu163位点、人USP9X的Phe2130位点周围序列的磷酸化肽段免疫动物获得。所有多克隆抗体经Protein A和肽亲和层析纯化。 本试剂盒中所提供的单克隆抗体是分别通过合成对应于人HAUSP蛋白羧基端序列的肽段或表达对应于人A20/TNFAIP3蛋白氨基端序列的重组蛋白并免疫动物获得。A20/TNFAIP3 (D13H3)兔单抗是与马克思普朗克生化研究所的Marc Schmidt-Supprian博士合作制备。 Western blot analysis of extracts from various cell lines using UCHL1 Antibody #3524. 使用UCHL1抗体#3524对不同细胞裂解物进行免疫印迹分析的结果。 Western blot analysis of extracts from various cell lines using UCHL3 Antibody #3525. 使用UCHL3抗体#3525对不同细胞裂解物进行免疫印迹分析的结果。 Western blot analysis of extracts from various cell lines using CYLD Antibody #4495. 使用CYLD抗体#4495对不同细胞裂解物进行免疫印迹分析的结果。 Western blot analysis of extracts from 293 cells, untreated (-) or treated (+) with hTNF-α #8902 (20 ng/ml for 20 minutes), using Phospho-CYLD (Ser418) Antibody #4500 (upper) or CYLD Antibody #4495 (lower). 使用磷酸化CYLD(Ser418)抗体#4500(上)或CYLD抗体#4495(下)对未经处理或经hTNF-α #8902 (20 ng/ml,20 minutes)处理的293细胞裂解物进行免疫印迹分析的结果。 Western blot analysis of extracts from various cell lines using HAUSP (D17C6) XP® Rabbit mAb #4833. 使用HAUSP (D17C6)XP® 兔单抗#4833对不同细胞裂解物进行免疫印迹分析的结果。 Western blot analysis of extracts from various cell lines using STAMBP Antibody #5245. 使用STAMBP抗体#5245对不同细胞裂解物进行免疫印迹分析的结果。 Western blot analysis of extracts from various cell lines using USP10 Antibody #5553. 使用USP10抗体#5553对不同细胞裂解物进行免疫印迹分析的结果。 Western blot analysis of extracts from various cell lines using A20/TNFAIP3 (D13H3) Rabbit mAb #5630. 使用A20/TNFAIP3 (D13H3)兔单抗#5630对不同细胞裂解物进行免疫印迹分析的结果。 Western blot analysis of extracts from various cell lines using USP9X Antibody #5751. 使用USP9X抗体#5751对不同细胞裂解物进行免疫印迹分析的结果。

Description

The DUB Antibody Sampler Kit提供对精选的去泛素化酶进行鉴定和检测的经济实用抗体组合。这个试剂盒内提供进行至少4次western blot检测的足够磷酸化抗体。

Western Blotting

Western Blotting

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.

IF-IC

IF-IC

Confocal immunofluorescent analysis of MCF7 cells using HAUSP (D17C6) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using HAUSP (D17C6) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, untreated or treated with hTNF-α #8902 (20 ng/ml for 20 minutes), using Phospho-CYLD (Ser418) Antibody (upper) or CYLD Antibody #4495 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using STAMBP Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from G-361, JJN-3, and A20 cell lines using A20/TNFAIP3 (D13H3) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated or treated with TPA #4174 (40 nM) and A23187 (2 μM) for various amounts of time, using A20/TNFAIP3 (D13H3) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS-7 cells, mock transfected (-) or transfected with a Myc-DDK-tagged human A20/TNFAIP3 construct (+), using A20/TNFAIP3 (D13H3) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using USP9X Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, untreated (-) or treated (+) with hTNF-α #8902 (20 ng/ml for 20 minutes), using Phospho-CYLD (Ser418) Antibody #4500 (upper) or CYLD Antibody #4495 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using STAMBP Antibody #5245.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using A20/TNFAIP3 (D13H3) Rabbit mAb #5630.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using UCHL1 Antibody #3524.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using HAUSP (D17C6) XP® Rabbit mAb #4833.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using USP9X Antibody #5751.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using UCHL3 (D25E6) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock-transfected (-) or transfected with a Myc/DDK-tagged cDNA expression construct encoding full-length human UCHL3 (hUCHL3, +), using UCHL3 (D25E6) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using CYLD (D1A10) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HCT 116 cells using USP10 (D7A5) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using USP10 (D7A5) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with a Myc/DDK-tagged cDNA expression construct encoding full-length human USP10 (hUSP10-Myc/DDK, +), using USP10 (D7A5) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using CYLD (D1A10) Rabbit mAb #8462.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using UCHL3 (D25E6) Rabbit mAb #8141.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using USP10 (D7A5) Rabbit mAb #8501.

IF-IC

IF-IC

Confocal immunofluorescent analysis of DU 145 (positive; left) and LNCaP (negative; right) cells using UCHL1 (D3T2E) XP® Rabbit mAb (green). Blue pseudocolor= DRAQ5® #4084 (fluorescent DNA dye).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded DU 145 (left; positive) and LNCaP (right; negative) cell pellets using UCHL1 (D3T2E) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using UCHL1 (D3T2E) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using UCHL1 (D3T2E) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded colorectal adenocarcinoma using UCHL1 (D3T2E) XP® Rabbit mAb in the presence of control peptide (left) or antigen specific peptide (right).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using UCHL1 (D3T2E) XP® Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human UCHL1 (hUCHL1-Myc/DDK; +), Myc/DDK-tagged full-length human UCHL3 (hUCHL3-Myc/DDK; +), Myc/DDK-tagged full-length human UCHL5 (hUCHL5-Myc/DDK; +), and Myc/DDK-tagged full-length human BAP1 (hBAP1-Myc/DDK; +), using UCHL1 (D3T2E) XP® Rabbit mAb (upper) or DYKDDDDK Tag Antibody #2368 (lower).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of K-562 cells (blue) and Ramos cells (green) using UCHL1 (D3T2E) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from Ramos and K-562 cells using UCHL1 (D3T2E) XP® Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). As expected, UCHL1 protein expression is not detected in K-562 cells.

Background

泛素酶(UBEs)催化蛋白质泛素化,去泛素化酶(DUB)在参与相反的过程——去泛素化(1,2)。 CYLD 去泛素化酶调节炎症和细胞增殖,通过去除几个NF-κB信号通路相关蛋白的泛素链下调NF-κB信号通路(3,4)。CYLD Ser418位的磷酸化降低CYLD去泛素化酶的活性,这对于IKKε驱动的转化过程具有重要作用(5)。 STAM-结合蛋白(STAMBP或AMSH)是一种胞内去泛素化酶,具有较强的针对K63位偶联泛素链的泛素异肽酶活性。A20的氨基端包含针对K63位偶联泛素链(如在TRAF6和RIP蛋白上的泛素化修饰)的去泛素活性,,而羧基末端包含针对K48位的泛素连接酶活性,并导致靶蛋白的降解(8)。这两种酶活性都在神经退行性疾病中发挥作用(9-11),调节神经元功能和精子发育(10,13,14)。 UCHL1结合单泛素,而UCHL3结合单泛素和泛素样分子NEDD8(11,12)。 HAUSP能够结合p53并对p53和相关调节蛋白MDM2去泛素化,从而增加这两种蛋白的稳定性(15,16)。 HAUSP的还调节其他泛素化的蛋白质,如叉头转录因子FOXO家族和有丝分裂的应力检查点蛋白CHFR(17,18)。 USP10似乎通过蛋白质相互作用和磷酸化被调节。USP10与RAS-gap SH3结构域结合蛋白(G3BP)相互作用,抑制其拆解泛素链的能力(19)。 ATM介导Thr42和Ser337位的磷酸化能够增加USP10的稳定性,促进其从细胞质再分配到细胞核,它在p53感应基因毒性应激反应后的去泛素化、稳定和激活过程中发挥作用(20)。 USP9X具有保守的具有半胱氨酸肽酶活性的催化结构域,可裂解泛素和多泛素结合物。虽然USP9X表达已被证明对于正常的哺乳动物发育(21-23)至关重要,但其机理还有待阐明。

Application References

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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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