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#64459 Autophagy Atg8 Family 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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キット内容 容量 用途 種交差性 ホモロジー† 検出分子量 アイソタイプ
LC3A (D50G8) XP® Rabbit mAb #4599 20 µl WB, IHC-P H, M, R Mk, Dg 14, 16 Rabbit IgG
LC3B (D11) XP® Rabbit mAb #3868 20 µl WB, IP, IHC-P, IF-IC, F H, M, R Mk, B, Pg 14, 16 Rabbit IgG
LC3C (D3O6P) Rabbit mAb #14736 20 µl WB H 14 Rabbit IgG
GABARAP (E1J4E) Rabbit mAb #13733 20 µl WB, IF-IC H, M, R 14, 16 Rabbit IgG
GABARAPL1 (D5R9Y) XP® Rabbit mAb #26632 20 µl WB, IF-IC, F H, M, R 14, 16 Rabbit IgG
GABARAPL2 (D1W9T) Rabbit mAb #14256 20 µl WB H, M, R, Mk 14 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl WB Goat

†Species predicted to react based on 100% sequence homology.

Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey

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 HeLa and NIH/3T3 cells, untreated or chloroquine-treated (50 μM, overnight), using LC3A (D50G8) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines, untreated (-) or treated overnight with chloroquine (50 μM) (+), using LC3B (D11) XP® Rabbit mAb (upper) or LC3B Antibody #2775 (lower).


Western Blotting

Western Blotting

Western blot analysis of A172 and C2C12 cells, untreated (-) or chloroquine-treated (50 μM, overnight; +), using GABARAP (E1J4E) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® GABARAPL2 siRNA I # 14239 (+) or SignalSilence® II GABARAPL2 siRNA II #14246 (+), using GABARAPL2 (D1W9T) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The GABARAPL2 (D1W9T) Rabbit mAb confirms silencing of GABARAPL2, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.

Western Blotting

Western Blotting

Western blot analysis of extracts from A172 and HOP-92 cells, untreated (-) or chloroquine-treated (50 μM, overnight; +), using LC3C (D3O6P) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (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 GABARAP protein (hGABARAP-Myc/DDK; +), human GABARAPL1 protein (hGABARAPL1-Myc/DDK; +), or human GABARAPL2 protein (hGABARAPL2-Myc/DDK; +) using GABARAPL1 (D5R9Y) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human glioblastoma multiforme using LC3A (D50G8) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® LC3B siRNA I #6212 (+) or SignalSilence® LC3B siRNA II #6213 (+), using LC3B (D11) XP® Rabbit mAb #3868 and α-Tubulin (11H10) Rabbit mAb #2125. The LC3B (D11) XP® Rabbit mAb confirms silencing of LC3B expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of LC3B siRNA.


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 GABARAP (hGABARAP-Myc/DDK; +), human GABARAPL1 (hGABARAPL1-Myc/DDK; +), or human GABARAPL2 (hGABARAPL2-Myc/DDK; +), using GABARAP (E1J4E) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using GABARAPL2 (D1W9T) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing full-length human LC3A protein (hLC3A; +), human LC3B protein (hLC3B; +), or human LC3C protein (hLC3C; +), using LC3C (D3O6P) Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, C2C12, and C6 cells, untreated (-) or treated with chloroquine (50 μM, overnight; +), using GABARAPL1 (D5R9Y) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human astrocytoma using LC3B (D11) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using LC3A (D50G8) XP® Rabbit mAb (left) or

Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).


IF-IC

IF-IC

Confocal immunofluorescent analysis of A172 cells, untreated (left) and chloroquine-treated (50 μM, overnight; right), using GABARAP (E1J4E) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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 GABARAP (hGABARAP-Myc/DDK; +), human GABARAPL1 (hGABARAPL1-Myc/DDK; +), or human GABARAPL2 (hGABARAPL2-Myc/DDK; +), using GABARAPL2 (D1W9T) Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells using GABARAPL1 (D5R9Y) XP® 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.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, control (left) or chloroquine-treated (right), using LC3B (D11) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, control (left) or choloroquine-treated (right), using LC3A (D50G8) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from A172 and HeLa cells, untreated (-) or treated overnight with chloroquine (50 μM) (+), using GABARAPL2 (D1W9T) Rabbit mAb (upper) or β-Actin (D6A8) #8457 (lower).


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with chloroquine (50 μM, 24 hr; right), using GABARAPL1 (D5R9Y) XP® Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells using LC3B (D11) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HCT-116 cells, untreated (left) or choroquine-treated (50 uM, overnight; right) using LC3B (D11) XP® Rabbit mAb (green) and β-Catenin (L54E2) Mouse mAb (Alexa Fluor® 555 Conjugate) #5612 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


バックグラウンド

Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents (1,2). Autophagy is generally activated by conditions of nutrient deprivation, but it has also been associated with a number of physiological processes, including development, differentiation, neurodegenerative diseases, infection, and cancer (3).

Atg8 is a ubiquitin-like protein that is critical for autophagosome formation. Atg8 is synthesized as a precursor protein that is processed by the cysteine protease Atg4, followed by lipidation with phosphatidylethanolamine (PE) in a ubiqutin-like conjugation pathway involving Atg7 and Atg3 (4). This processing of Atg8, which is described as a conversion from type-I to type-II forms, is frequently described as a marker for autophagy. The type-II form of Atg8 is incorporated into maturing autophagosomes and leads to the recruitment of additional autophagy components, including cargo receptors like SQSTM1/p62. While yeast has a single Atg8 gene, many eukaryotes have at least six orthologs, including three microtubule-associated protein 1 light chain 3 (MAP1LC3/LC3) family members (LC3A, LC3B, and LC3C) and three GABAA receptor associated protein (GABARAP) family members (GABARAP, GABARAPL1/GEC1, and GABARAPL2/GATE-16). While highly conserved, these various family members can have important differences in their post-translational processing, expression profile, and protein interactions including distinct cargo receptor. This complexity within the Atg8 family is critical for selective mechanisms of autophagy that have been reported (5, 6).

使用例
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14256   GABARAPL2 (D1W9T) Rabbit mAb
14736   LC3C (D3O6P) Rabbit mAb
26632   GABARAPL1 (D5R9Y) XP® Rabbit mAb
3868   LC3B (D11) XP® Rabbit mAb
4599   LC3A (D50G8) XP® Rabbit mAb
6883   SignalFire™ ECL Reagent
7074   Anti-rabbit IgG, HRP-linked Antibody
7727   Biotinylated Protein Ladder Detection Pack

DRAQ5 is a registered trademark of Biostatus Limited.
Tween is a registered trademark of ICI Americas, Inc.
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.

本製品は試験研究用です。

Autophagy Atg8 Family Antibody Sampler Kit

Metabolic Reprogramming in Disease

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