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#89947 ER Stress-induced Autophagy Antibody Sampler Kit

 
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希望納入価格 (円)
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2019年1月17日15時10分 現在
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#89947T1 Kit99,000
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キット内容 容量 用途 種交差性 検出分子量 アイソタイプ
BiP (C50B12) Rabbit mAb #3177 20 µl WB, IHC-P, IHC-F, F H, M 78 Rabbit IgG
eIF2α (D7D3) XP® Rabbit mAb #5324 20 µl WB, IP, IHC-P H, M, R, Mk 38 Rabbit IgG
Phospho-eIF2α (Ser51) (D9G8) XP® Rabbit mAb #3398 20 µl WB, IP, IHC-P H, M, R, Mk, Dm 38 Rabbit IgG
Atg12 (D88H11) Rabbit mAb #4180 20 µl WB, IP H, M, R, Mk 16, 55 Rabbit IgG
Beclin-1 (D40C5) Rabbit mAb #3495 20 µl WB, IP H, M, R, Mk 60 Rabbit IgG
JNK1 (2C6) Mouse mAb #3708 20 µl WB H, M, R, Mk 46, 54 Mouse IgG1
Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb #4668 20 µl WB, IP, IHC-P H, M, R, Dm, Sc 46, 54 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl WB Goat
Anti-mouse IgG, HRP-linked Antibody #7076 100 µl WB Horse

Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), IHC-F=Immunohistochemistry (Frozen), F=Flow Cytometry, IP=Immunoprecipitation
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey, Dm=D. melanogaster, Sc=S. cerevisiae

貯法
-20℃
社内データ

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 extracts from various cell lines using BiP (C50B12) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, untreated or UV-treated, NIH/3T3 cells, untreated or UV-treated and C6 cells, untreated or anisomycin-treated, using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from C2C12 cells, untreated or thapsigargin-treated, using Phospho-eIF2α (Ser51) (D9G8) XP® Rabbit mAb (upper) or eIF2α Antibody #9722 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Beclin-1 (D40C5) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of HeLa cell extracts, untransfected (lane 1), mock-transfected (lane 2) or transfected with SignalSilence® SAPK/JNK siRNA I #6232 (lane 3) or SignalSilence® SAPK/JNK siRNA II #6233 (lane 4) for 72 hours, using JNK1 (2C6) Mouse mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Atg12 (D88H11) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using eIF2α (D7D3) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded 293T cells untreated (left) or UV-treated (right) using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human glioblastoma using BiP (C50B12) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, untreated (left) or λ phosphatase-treated (right), using Phopsho-eIF2α (Ser51) (D9G8) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from indicated cell lines, untreated or UV-treated (40 J/m2, 30 min recovery), using JNK1 (2C6) Mouse mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® Beclin-1 siRNA I #6222 (+) or SignalSilence® Beclin-1 siRNA II (+), using Beclin-1 (D40C5) XP® Rabbit mAb #3495 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The Beclin-1 (D40C5) XP® Rabbit mAb confirms silencing of Beclin-1 expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of Beclin-1 siRNA.

IP

IP

Immunoprecipitation/western blot analysis of lysates from HeLa cells. Lane 1 contains lysate input (10%), lane 2 was immunoprecipitated with non-specific rabbit IgG, lane 3 was immunoprecipitated with eIF2α (D7D3) XP® Rabbit mAb #5324. Western blot analysis was performed using eIF2α (L57A5) Mouse mAb #2103.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb in the presence of control peptide (left) or Phospho-SAPK/JNK (Thr183/Tyr185) Blocking Peptide #1215 (right).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using BiP (C50B12) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-eIF2α (Ser51) (D9G8) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse colon using eIF2α (D7D3) XP® Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using BiP (C50B12) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lymphoma using Phospho-eIF2α (Ser51) (D9G8) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using eIF2α (D7D3) XP® Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using BiP (C50B12) Rabbit mAb in the presence of control peptide (left) or BiP Blocking Peptide #1084 (right).

IHC-F (frozen)

IHC-F (frozen)

Immunohistochemical analysis of frozen SKOV-3 xenograft using BiP (C50B12) Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of A204 cells using BiP (C50B12) Rabbit mAb (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.


バックグラウンド

The endoplasmic reticulum (ER) is an organelle with essential biosynthetic and signaling functions in eukaryotic cells (1). Post synthesis of secretory and transmembrane proteins on polysomes, proteins are translocated into the ER where they are often modified by disulfide bond formation, amino-linked glycosylation, and folding. Different physiological and pathological conditions can disturb proper protein folding in the ER causing ER stress (1). ER stress activates an intracellular signaling transduction pathway called unfolded protein response (UPR) and autophagy to avoid cell death (2). The main role of UPR is to improve the protein load on the ER by shutting down protein translation and gene transcription to enhance ER's folding capacity (2). On the other hand, autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmc contents (3,4). One of the chaperones aiding in proper protein folding is Binding immunoglobulin Protein (BiP) (5,6). BiP works by binding to misfolded proteins to prevent them from forming aggregates and assists in proper refolding (7). The molecular machinery of autophagy was largely discovered in yeast and referred to as autophagy-related (Atg) genes. Formation of the autophagosome involves a ubiquitin-like conjugation system in which Atg12 is covalently bound to Atg5 and targeted to autophagosome vesicles (8-10). One of the proteins critical to autophagy process is Beclin-1, the mammalian orthologue of the yeast autophagy protein Apg6/Vps30 (11). Beclin-1 can complement defects in yeast autophagy caused by loss of Apg6 and can also stimulate autophagy when overexpressed in mammalian cells (12). Mammalian Beclin-1 was originally isolated in a yeast two-hybrid screen for Bcl-2 interacting proteins and has been shown to interact with Bcl-2 and Bcl-xL, but not with Bax or Bak (13). Phosphorylation of the eukaryotic initiation factor 2 (eIF2) α subunit is a well-documented mechanism to downregulate protein synthesis under a variety of stress conditions. eIF2 binds GTP and Met-tRNAi and transfers Met-tRNA to the 40S subunit to form the 43S preinitiation complex (14,15). Kinases that are activated by viral infection (PKR) can phosphorylate the α subunit of eIF2 (16,17). Induction of PKR by IFN-γ and TNF-α induces potent phosphorylation of eIF2α at Ser51 (18,19). There are three SAPK/JNK genes each of which undergoes alternative splicing, resulting in numerous isoforms (20). The IRE1, a transmembrane serine/threonine kinase (21,22), through its kinase activity activates SAPK/JNK in the early stage of ER stress in order to induce autophagosome formation (23).

使用例
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関連製品
12630   SignalFire™ Plus ECL Reagent
12757   SignalFire™ Elite ECL Reagent
3177   BiP (C50B12) Rabbit mAb
3398   Phospho-eIF2α (Ser51) (D9G8) XP® Rabbit mAb
3495   Beclin-1 (D40C5) Rabbit mAb
3708   JNK1 (2C6) Mouse mAb
4180   Atg12 (D88H11) Rabbit mAb
4668   Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb
5324   eIF2α (D7D3) XP® Rabbit mAb
6883   SignalFire™ ECL Reagent
7074   Anti-rabbit IgG, HRP-linked Antibody
7076   Anti-mouse IgG, HRP-linked Antibody
7727   Biotinylated Protein Ladder Detection Pack
9997   Tris Buffered Saline with Tween® 20 (TBST-10X)
9999   Nonfat Dry Milk

XP is a registered trademark of Cell Signaling Technology, Inc.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

本製品は試験研究用です。

ER Stress-induced Autophagy Antibody Sampler Kit

Immune Cell Signaling Pathways

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