Cell Signaling Technology

Product Pathways - Protein Folding

Protein Folding and Stability Antibody Sampler Kit #4889

developmentally down-regulated 8   E2   EMC19   F-box protein Skp2   F-box/LRR-repeat protein 1   FBL1   FBXL1   FLB1   GAP modifying protein 1   general   GMP1   HSMT3   Interferon-induced 15 kDa protein   motif   Nedd-8   Neddylin   neural precursor cell expressed   OCP-2   OCP-II protein   OCP2   Organ of Corti protein 2   p19   p19A   p19skp1   p45skp2   RNA polymerase II elongation factor-like protein   S-phase kinase-associated protein 1A   S-phase kinase-associated protein 2   SCF Complex   Sentrin 2   Sentrin 3   SIII   SKP1A   SMT 3B   SUMO-2/3   SUMO1   TCEB1L   Transcription elongation factor B   ub   UBC   Ubiquitin carrier protein   Ubiquitin cross-reactive protein (UCRP)   Ubiquitin protein ligase   Ubiquitin-conjugating enzyme E2-32 kD   ubiquitin-like protein NEDD8   Ubiquitin-like protein SMT3C   Ubiquitin-like protein UBL1   Ubiquitin-related protein SUMO  

REACTIVITY

No. Size Price
4889T 1 Kit ( 8 x 20 µl ) ¥6,597.00 现货查询 购买询价
Kit Includes Quantity Applications Reactivity Homology† MW (kDa) Isotype
Skp1 Antibody #2156 20 µl W,IF-IC,F, H,M,R, 19 Rabbit
Skp2 (D3G5) XP® Rabbit mAb #2652 20 µl W,IP,IHC-P,IF-IC, H,Mk, 48 Rabbit IgG
NEDD8 (19E3) Rabbit mAb #2754 20 µl W,IP,IHC-P,F, H,M,R,Mk, X,Z,B, 9 Rabbit IgG
ISG15 (22D2) Rabbit mAb #2758 20 µl W,IP, H, 15 Rabbit IgG
Ubiquitin (P4D1) Mouse mAb #3936 20 µl W,IHC-P, All, Mouse IgG1
SUMO-1 Antibody #4930 20 µl W,IP,IHC-P,IF-IC, H,M,R,Mk, Rabbit
SUMO-2/3 (18H8) Rabbit mAb #4971 20 µl W,IF-IC, H,M,R, Rabbit IgG
UBC3 Antibody #4997 20 µl W,F, H,M,R, 32 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl W, Goat
Anti-mouse IgG, HRP-linked Antibody #7076 100 µl W, Horse

Specificity / Sensitivity

Skp1抗体识别内源性的Skp1总蛋白。Skp2抗体识别内源性Skp2总蛋白(α, β and γ 亚型),与其他SKP蛋白无交叉反应。ISG15 (22D2)兔单抗识别的内源性ISG15未切割前体的表达蛋白,这一抗体不识别激活型(切割后)或偶联到目标蛋白上的ISG15,这一抗体不与其他泛素家族成员包括泛素、SUMO-1、2、3和NEDD8发生交叉反应。NEDD8 (19E3)兔单抗可以检测游离的和偶联后的NEDD8蛋白,这一抗体不与其他泛素家族成员包括泛素、SUMO-1、2、3和ISG15发生交叉反应。Ubiquitin (P4D1)鼠单抗可以检测泛素、多泛素和泛素化蛋白。UBC3抗体识别内源性UBC3和UBC3B总蛋白水平。SUMO-1抗体检测重组SUMO-1和内源性SUMO化蛋白质水平(如SUMO-1-RanGAP, 90kD)。SUMO-2/3抗体检测内源性SUMO-2/3总蛋白水平,但与重组SUMO-1不发生交叉反应。

Source / Purification

上述多克隆抗体SUMO-1(#4930)、UBC3(#4997)、Skp2(#4358)和Skp1(#2156)是通过合成与人SUMO-1、UBC3、Skp2和Skp1蛋白氨基酸序列一致的肽段并免疫动物获得,抗体经Protein A和肽亲和层析纯化。 SUMO-2/3 (18H8)兔单抗(#4971)是通过合成与SUMO-3蛋白氨基末端的氨基酸残基序列一致的肽段,并免疫动物获得。Ubiquitin (P4D1)鼠单抗(#3936)是通过合成1-76全长牛泛素肽段并免疫动物获得。ISG15 (22D2)兔单抗是通过合成与人ISG15蛋白氨基末端序列一致的肽段免疫动物获得。NEDD8 (19E3)兔单抗是通过合成与人NEDD8蛋白氨基末一致端序列的肽段免疫动物获得。 Western blot analysis of extracts from PC12 and NIH/3T3 cells using Skp1 Antibody #2156. 使用Skp1抗体#2156对PC12和NIH/3T3细胞裂解物进行免疫印迹分析的结果。 Western blot analysis of lysates from HeLa, RAW, C6 and COS cells, using NEDD8 (19E3) Rabbit mAb #2754. 使用NEDD8 (19E3)兔单抗#2754对HeLa、RAW、C6和COS细胞裂解物进行免疫印迹分析的结果。 Western blot analysis of lysates from HeLa and A549 cells, untreated (-) or treated with IFN-α (1000 U/mL) (+) for 24 hours, using ISG15 (22D2) Rabbit mAb #2758. 使用ISG15(22D2)兔单克隆抗体#2758对未经处理(-)或经α干扰素(1000 U / ml的,24小时)处理(+)的HeLa和A549细胞裂解物进行免疫印迹分析的结果。 Western blot analysis of extracts from 293 and HeLa cells, untreated or MG-132 (a 26S proteosome inhibitor) treated (50 µM for 90 minutes), using Ubiquitin (P4D1) Mouse mAb #3936. 使用泛素(P4D1)小鼠单克隆抗体#3936对未经处理或经MG-132(26S蛋白酶抑制剂)处理(50 µM,90 分钟)的293和HeLa细胞提取物进行Western blot分析的结果。 Western blot analysis of extracts from HeLa, K562 and COS cells using Skp2 Antibody #4358. 使用Skp2抗体#4358对HeLa、K562和COS细胞裂解物进行免疫印迹分析的结果。 Western blot analysis of recombinant GST-SUMO-1 protein (38 kDa) and extracts from CAD, HeLa and PC-12 cells using SUMO-1 Antibody #4930. 使用SUMO-1抗体#4930对重组GST-SUMO-1蛋白质(38 kDa)和CAD、HeLa和PC-12细胞提取物进行Western blot分析的结果。 Western blot analysis of recombinant GST-SUMO-1 protein (38 kDa), recombinant SUMO-2, recombinant SUMO-3, and extracts from C6 and PC12 cells, using SUMO-2/3 (18H8) Rabbit mAb #4971 (upper) and SUMO-1 Antibody #4930 (lower). 使用SUMO-2/3(18H8)兔单克隆抗体#4971(上)和SUMO-1抗体#4930(下)丢重组GST-SUMO-1蛋白质(38 kDa)、重组SUMO-2、重组SUMO-3、C6和PC12细胞提取物进行免疫印迹分析的结果。 Western blot analysis of extracts from Jurkat, CAD and C6 cells using UBC3 Antibody #4997. 使用UBC3抗体#4997对Jurkat、CAD和C6细胞裂解物进行免疫印迹分析的结果。

Description

这一抗体小包装试剂盒是用于研究蛋白折叠和稳定性过程相关蛋白的经济适用组合,这一试剂盒包含有每个目标蛋白足够进行4次western blot检测的一抗和二抗。

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.

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.

Western Blotting

Western Blotting

Western blot analysis of 293 and HeLa cells, untreated or treated with the 26S proteasome inhibitor MG132 (50 µM, 90 minutes), using Ubiquitin (P4D1) Mouse mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded breast carcinoma showing nuclear and cytoplasmic ubiquitin localization, using Ubiquitin (P4D1) Mouse mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of untreated Jurkat cells using UBC3 antibody (blue) compared to a nonspecific negative control antibody (red).

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat, CAD and C6 cells using UBC3 Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from PC12 and NIH/3T3 cells, using Skp1 Antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of NIH/3T3 cells using Skp1 antibody (blue) compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal immunofluorescent images of NIH/3T3 cells labeled with Skp1 Antibody (green, left) compared to an isotype control (right). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Western Blotting

Western Blotting

Western blot analysis of lysates from HeLa and A549 cells, untreated (-) or treated with IFN-α (1000 U/mL) (+) for 24 hours, using ISG15 (22D2) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of ubiquitin, NEDD8, ISG15 and SUMO-2/3 recombinant proteins (5 ng each), using NEDD8 (19E3) Rabbit mAb, Ubiquitin (P4D1) Mouse mAb #3936, ISG15 Antibody #2743 and SUMO-2/3 Antibody #4974.

Western Blotting

Western Blotting

Western blot analysis of lysates from HeLa, RAW, C6 and COS cells, using NEDD8 (19E3) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of recombinant GST-SUMO-1 protein (38 kDa) and extracts from CAD, HeLa and PC-12 cells, using SUMO-1 Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using NEDD8 (19E3) Rabbit mAb, preincubated with control peptide (left) or Nedd8 Blocking Peptide #1048 (right).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of untreated HeLa cells using NEDD8 (19E3) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, using SUMO-1 Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 (red).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, usinig SUMO-1 Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using SUMO-1 Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using SUMO-1 Antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of SH-SY5Y cells labeled with SUMO-2/3 (18H8) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma, using NEDD8 (19E3) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using NEDD8 (19E3) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of recombinant GST-SUMO-1 protein (38 kDa), recombinant SUMO-2, recombinant SUMO-3, and extracts from C6 and PC12 cells, using SUMO-2/3 (18H8) Rabbit mAb (upper) and SUMO-1 Antibody #4930 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 and HeLa cells, untreated or MG-132 (a 26S proteosome inhibitor) treated (50 µM for 90 minutes), using Ubiquitin (P4D1) Mouse mAb #3936.

Western Blotting

Western Blotting

Western blot analysis of recombinant GST-SUMO-1 protein (38 kDa) and extracts from CAD, HeLa and PC-12 cells using SUMO-1 Antibody #4930.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat, CAD and C6 cells using UBC3 Antibody #4997.

Western Blotting

Western Blotting

Western blot analysis of extracts from PC12 and NIH/3T3 cells using Skp1 Antibody #2156.

Western Blotting

Western Blotting

Western blot analysis of recombinant GST-SUMO-1 protein (38 kDa), recombinant SUMO-2, recombinant SUMO-3, and extracts from C6 and PC12 cells, using SUMO-2/3 (18H8) Rabbit mAb #4971 (upper) and SUMO-1 Antibody #4930 (lower).

Western Blotting

Western Blotting

Western blot analysis of lysates from HeLa and A549 cells, untreated (-) or treated with IFN-α (1000 U/mL) (+) for 24 hours, using ISG15 (22D2) Rabbit mAb #2758.

Western Blotting

Western Blotting

Western blot analysis of lysates from HeLa, RAW, C6 and COS cells, using NEDD8 (19E3) Rabbit mAb #2754.

IF-IC

IF-IC

Confocal immunofluorescent analyis of HeLa cells expressing either a non-targeting shRNA (shNT; left) or shSkp2 (right) using Skp2 (D3G5) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red) (Cell lines kindly provided by Dr. Wenyi Wei, Harvard Medical School.)

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using Skp2 (D3G5) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Skp2 (D3G5) XP® 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 Skp2 (hSkp2-Myc/DDK, +), using Skp2 (D3G5) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells expressing either non-targeting shRNA (shNT) or shSkp2, using Skp2 (D3G5) XP® Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lymphoma using Skp2 (D3G5) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Skp2 (D3G5) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Skp2 (D3G5) XP® Rabbit mAb #2652.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using SUMO-2/3 (18H8) Rabbit mAb (green). Actin filaments were labeled with DyLight® 554 Phalloidin #13054 (red).

Background

小调节蛋白泛素往往通过共价方式连接到许多细胞蛋白质,这一标签介导标记蛋白进入蛋白酶体降解。泛素首先与活化酶E1形成硫酯键复合体而被激活,然后被转移到泛素载体蛋白E2,然后由E2转移到E3并连接到靶蛋白赖氨酸的epsilon位氨基上(1),泛素——蛋白酶体途径参与多种生物过程和疾病发生(2-3)。泛素样蛋白家族包含三个小泛素相关修饰蛋白SUMO-1,-2和-3,神经前体细胞表达发育下调蛋白8(NEDD8)和干扰素刺激的15kDa蛋白(ISG15) (4-6)。他们共价连接到靶蛋白是可逆的多步骤的过程,类似于蛋白泛素化过程。成熟的修饰分子与激活酶E1偶联,然后结合到E2,然后通过E3连接酶结合到靶蛋白(7-10)。泛素主要调节目标蛋白质的降解(8),而SUMO、NEDD8和ISG15调控一部分底物蛋白以调节其他各种生物过程(4,11-18)。泛素化过程中,E2和E3蛋白的不同组合和相互作用产生可变的底物特异性(4)。 UBC3和UBC3B都属于E2泛素载体蛋白(19,20)。 SCF(Skp1/CUL1/F-box)E3泛素连接酶蛋白复合体是由三种蛋白质组分,包括S期激酶相关蛋白1(SKP1),cullin同源1(CUL1)和Skp2的F-box蛋白(21 -23)。

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

U.S. Patent No. 5,675,063.

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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