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#43110 Mitophagy Antibody Sampler Kit

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2019年3月20日11時40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
SQSTM1/p62 (D5E2) Rabbit mAb #8025 20 µl WB, IP H, Mk 62 Rabbit IgG
NDP52 (D1E4A) Rabbit mAb #60732 20 µl WB H 52, 60 Rabbit IgG
Optineurin (D2L8S) Rabbit mAb #58981 20 µl WB H 75 Rabbit IgG
Parkin (Prk8) Mouse mAb #4211 20 µl WB, IP H, M, R 50 Mouse IgG2b
PINK1 (D8G3) Rabbit mAb #6946 20 µl WB, IP H 60, 50 Rabbit IgG
BNIP3 (D7U1T) Rabbit mAb #44060 20 µl WB, IP, IHC-P, IF-IC H 22-28, 50-55 Rabbit IgG
BNIP3L/Nix (D4R4B) Rabbit mAb #12396 20 µl WB, IP, IF-IC H, M, R, Mk 38, 76 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
Phospho-Ubiquitin (Ser65) (E2J6T) Rabbit mAb #62802 20 µl WB, IP H M, R 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, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Reactivity Key: H=Human, Mk=Monkey, M=Mouse, R=Rat

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 PC12 cells, fetal rat brain and mouse brain, using Parkin (Prk8) Mouse mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® SQSTM1/p62 siRNA I #6394 (+) or SignalSilence® SQSTM1/p62 siRNA II #6399 (+), using SQSTM1/p62 (D5E2) Rabbit mAb (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The SQSTM1/p62 (D5E2) Rabbit mAb confirms silencing of SQSTM1/p62 expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.


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 extracts from 293T cells, mock transfected (-) or transfected with a cDNA construct expressing full-length human PINK1 (hPINK1, +) using PINK1 (D8G3) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of A172 and HeLa cells, untreated (-) or cobalt chloride-treated (100 μM, overnight; +), using BNIP3L/Nix (D4R4B) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa or MCF7 cells, untreated (-) or cobalt chloride-treated (100 μM, overnight; +), using BNIP3 (D7U1T) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using NDP52 (D1E4A) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Optineurin (D2L8S) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from PC-3 cells, untreated (-) or treated with carbonyl cyanide 3-chlorophenylhydrazone (CCCP; 30 μM, 6 hr; +), using Phospho-Ubiquitin (Ser65) (E2J1T) Rabbit mAb (upper), PINK1 (D8G3) Rabbit mAb #6946 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using SQSTM1/p62 (D5E2) 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 HeLa cells, untreated (-) or treated with CCCP (10 μM, 24 hr; +), using PINK1 (D8G3) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using BNIP3L/Nix (D4R4B) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc-tagged full-length human BNIP3 protein (hBNIP3-Myc; +), using BNIP3 (D7U1T) Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from PC-3 cells treated with carbonyl cyanide 3-chlorophenylhydrazone (CCCP; 30 μM, 6 hr ). Lysates were treated with (+) or without (-) Lambda Phosphatase (λ-Phosphatase) and Calf Intestinal Phosphatase (CIP). Western blot was performed using Phospho-Ubiquitin (Ser65) (E2J1T) Rabbit mAb (upper) or 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).

Western Blotting

Western Blotting

Western blot analysis of extracts from SK-MEL-2 cells, untreated (-) or starved overnight in Earle's Balanced Salt Solution (EBSS) (+), using SQSTM1/p62 (D5E2) Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing DYKDDDDK-tagged full-length human BNIP3L/Nix (hBNIP3L/Nix-DYKDDDDK; +), using BNIP3L/Nix (D4R4B) Rabbit mAb. The BNIP3L/Nix construct was kindly provided by Dr. Reuben Shaw, Salk Institute for Biological Studies, La Jolla, CA.

IP

IP

Immunoprecipitation of BNIP3 from MCF7 cells treated with cobalt chloride (100 μM; overnight). Lane 1 represents 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is BNIP3 (D7U1T) Rabbit mAb. Western blot analysis was performed using BNIP3 (D7U1T) Rabbit mAb. A conformation-specific secondary antibody was used to avoid reactivity with IgG.

IP

IP

Immunoprecipitation of phospho-ubiquitin (Ser65) from extracts of PC-3 cells treated with carbonyl cyanide 3-chlorophenylhydrazone (CCCP; 30 μM, 6 hr). Lane 1 is 10% input, lane 2 is precipitated with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-Ubiquitin (Ser65) (E2J6T) Rabbit mAb. Western blot was performed using Phospho-Ubiquitin (Ser65) (E2J6T) Rabbit mAb. Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 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.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a tagged human SQSTM1/p62 construct (+), using SQSTM1/p62 (D5E2) Rabbit mAb.

IP

IP

Immunoprecipitation of BNIP3L/Nix from A172 cells, treated with cobalt chloride (100 μM, overnight), using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or BNIP3L/Nix (D4R4B) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using BNIP3L/Nix (D4R4B) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using BNIP3 (D7U1T) Rabbit mAb.

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 A172 cells, untreated (left) or cobalt chloride-treated (100 μM, overnight; right), using BNIP3L/Nix (D4R4B) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using BNIP3 (D7U1T) Rabbit mAb.

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).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded MCF7 cell pellets, untreated (left) or cobalt chloride-treated (right), using BNIP3 (D7U1T) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded normal human liver using BNIP3 (D7U1T) Rabbit mAb in the presence of control peptide (left) and antigen-specific peptide (right).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with cobalt chloride (100 μM, 24 hr; right), using BNIP3 (D7U1T) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalliodin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

バックグラウンド

Autophagy is a catabolic process for the autophagosome-lysosomal degradation of bulk cytoplasmic contents (1, 2). Selective autophagy targets the degradation of distinct sets of substrates and organelles (3-5). One of the best studied examples of selective autophagy involves the clearance of damaged mitochondria through a process called mitophagy. Several pathways have been described for various contexts of mitophagy, including the FUNDC1 pathway, the BNIP3 and BNIP3L/Nix pathway, and the PINK1/Parkin pathway. FUNDC1 is a mitochondrial protein that is phosphorylated by the autophagy kinase ULK1 and regulates hypoxia induced mitophagy (6, 7). BNIP3L/Nix and BNIP3 are members of the Bcl-2 family of apoptosis regulators that are expressed on mitochondria, induced by hypoxia, and have have been shown to play a role in mitophagy (8). BNIP3L/Nix is also important in the autophagic maturation of erythroid cells (9). FUNDC1, BNIP3 and BNIP3L/Nix bind to LC3 family members, targeting the mitochondria to the autophagosome.

Non-hypoxic induction of mitophagy can be regulated by the PINK1/Parkin pathway, which plays causative roles in neurodegenerative disease, most notably Parkinson’s disease (10, 11). PINK1 is a mitochondrial serine/threonine kinase that is stabilized on the outer mitochondrial membrane of damaged mitochondria. Substrates of PINK1 include the E3 ubiquitin ligase Parkin and ubiquitin itself (12-14). Phosphorylation of Parkin as well as binding to phosphorylated ubiquitin leads to accumulation of ubiquitinated chains on multiple mitochondrial proteins. Ubiquitinated proteins are recognized by selective cargo receptors including SQSTM1/p62, Optineurin, and NDP52 (15-16). Autophagy cargo receptors contain an LC3-interacting region (LIR) required for binding to Atg8/LC3 family members and targeting to the autophagosome (3).

The Mitophagy Antibody Sampler Kit provides an economical means of detecting proteins involved in the process of mitophagy. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

使用例
製品をご使用いただいて研究を発表されましたら、ぜひお知らせください
 
関連製品
12396   BNIP3L/Nix (D4R4B) Rabbit mAb
12630   SignalFire™ Plus ECL Reagent
12757   SignalFire™ Elite ECL Reagent
13953   Prestained Protein Marker, Broad Range (11-190 kDa)
3868   LC3B (D11) XP® Rabbit mAb
4211   Parkin (Prk8) Mouse mAb
44060   BNIP3 (D7U1T) Rabbit mAb
58981   Optineurin (D2L8S) Rabbit mAb
60732   NDP52 (D1E4A) Rabbit mAb
62802   Phospho-Ubiquitin (Ser65) (E2J6T) Rabbit mAb
6883   SignalFire™ ECL Reagent
6946   PINK1 (D8G3) Rabbit mAb
7072   Phototope-HRP Western Blot Detection System, Anti-mouse IgG, HRP-linked Antibody
7074   Anti-rabbit IgG, HRP-linked Antibody
7076   Anti-mouse IgG, HRP-linked Antibody
7727   Biotinylated Protein Ladder Detection Pack
8025   SQSTM1/p62 (D5E2) Rabbit mAb
9997   Tris Buffered Saline with Tween® 20 (TBST-10X)

Tween is a registered trademark of ICI Americas, Inc.
DRAQ5 is a registered trademark of Biostatus Limited.
SignalStain is a trademark of Cell Signaling Technology, 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.
DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.

本製品は試験研究用です。

Mitophagy Antibody Sampler Kit

Metabolic Reprogramming in Disease

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