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Home/Search results/Products/Arrays/Antibody Arrays/C-Series/Apoptosis Arrays/Mouse Apoptosis Signaling Pathway Array C1

Manual / SDS

NIH/3T3 cells were grown to 80% confluency and then serum starved overnight. Cells were treated (left panel) with Camptothecin (CPT) for 16 hours. Data shown are from a 20 second exposure using a chemiluminescence imaging system.

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Assay Format

Pathway:

Akt Signaling
HER/ErbB Signaling
HIF-1 alpha Signaling
IGF Signaling
JAK/STAT Signaling
MAPK Signaling
mTOR Signaling
NFkB Signaling
p53 Signaling
PI3K-AKT Signaling
PKC Signaling
TGF-beta Signaling

Number of Targets Detected:

Species Detected:

Mouse

Compatible Sample Types:

Cell Culture Supernatants
Cell Lysates
Plasma
Serum
Tissue Lysates

Design Principle:

Sandwich-based

Method of Detection:

Chemiluminescence

Quantitative/Semi-Quantitative:

Semi-Quantitative

Solid Support:

Membrane

Product Specifications

Size:

2, 4, or 8 Sample Kit

Protein Information

Gene Symbols:

Product Features

Easy to use
No specialized equipment needed
Compatible with nearly any liquid sample
Proven technology (many publications)
Highly sensitive (pg/ml)
Sandwich ELISA specificity
Higher density than ELISA, Western blot or bead-based multiplex

Target Names

Scroll over each target protein for more information
AKT (P-Ser473) Gene Symbol: Akt1 Akt Rac Gene ID: 11651 Accession #: P31750 ELISA Kit: RayBio Human/Mouse/Rat Phospho-Akt (S473) ELISA	ATM (P-Ser1981) Gene Symbol: Atm Gene ID: 11920 Accession #: Q62388 ELISA Kit: RayBio Human Phospho-ATM (S1981) ELISA	BAD (P-Ser112) Gene Symbol: Bad Bbc6 Gene ID: 12015 Accession #: Q61337 ELISA Kit: RayBio Human Phospho-BAD (S112) ELISA	Caspase-3 (P-Asp175) Gene Symbol: Casp3 Cpp32 Gene ID: 12367 Accession #: P70677 ELISA Kit: RayBio Human Cleaved-Caspase-3 (D175) and Caspase-3 ELISA	Caspase-7 (P-Asp198) Gene Symbol: Casp7 Lice2 Mch3 Gene ID: 12369 Accession #: P97864 ELISA Kit: RayBio Human/Mouse Cleaved-Caspase-7 (D198) and Caspase-7 ELISA
CHK1 (P-Ser296) Gene Symbol: Chek1 Chk1 Gene ID: 12649 Accession #: O35280 ELISA Kit: RayBio Human Phospho-Chk1 (S280) ELISA	eIF-2a (P-Ser52) Gene Symbol: Eif2s1 Eif2a Gene ID: 13665 Accession #: Q6ZWX6 ELISA Kit: RayBio Human Phospho-eIF-2A (S52) ELISA	ERK1/2 (P-Thr202) Accession #: Q63844 \| P63085 ELISA Kit: RayBio Human/Mouse/Rat Phospho-Erk1 (T202/Y204) + Phospho-Erk2 (Y185/Y187) ELISA	HSP27 (P-Ser82) Gene Symbol: Hspb1 Hsp25 Hsp27 Gene ID: 15507 Accession #: P14602 ELISA Kit: RayBio Human Phospho-HSP27 (S82) ELISA	IKBa (P-Ser32) Gene Symbol: Nfkbia Ikba Gene ID: 18035 Accession #: Q9Z1E3 ELISA Kit: RayBio Human Phospho-I-kappa B alpha (S32) ELISA
JNK (P-Thr183) Gene Symbol: Mapk8 Jnk1 Prkm8 Gene ID: 26419 Accession #: Q91Y86 ELISA Kit: RayBio Human/Mouse/Rat Phospho-JNK (T183/Y185) ELISA	NF-kB P65 (P-Ser536) Gene Symbol: Rela Nfkb3 Gene ID: 19697 Accession #: Q04207 ELISA Kit: RayBio Human Phospho-NF-KB p65 (S536) ELISA	p27 (P-Thr198) Gene Symbol: Cdkn1b Gene ID: 12576 Accession #: P46414 ELISA Kit: RayBio Human Phospho-P27/Kip1 (T198) ELISA	p38 (P-Thr180/Tyr182) Gene Symbol: Mapk14 Crk1 Csbp1 Csbp2 Gene ID: 26416 Accession #: P47811 ELISA Kit: RayBio Human/Mouse Phospho-p38 alpha (T180/Y182) ELISA	P53 (P-Ser15) Gene Symbol: Tp53 P53 Trp53 Gene ID: 22059 Accession #: P02340 ELISA Kit: RayBio Human/Mouse/Rat Phospho-P53 (S15) ELISA
SMAD2 (P-Ser245) Gene Symbol: Smad2 Madh2 Madr2 Gene ID: 17126 Accession #: Q62432 ELISA Kit: RayBio Human/Mouse/Rat Phospho-SMAD2 (SER245/250/255) ELISA	TAK1 (P-Ser412) Gene Symbol: Map3k7 Tak1 Gene ID: 26409; Accession #: Q62073 ELISA Kit: RayBio Human Phospho-TAK1 (Ser412) ELISA

Typical Results

Figure 1: NIH/3T3 cells were grown to 80% confluency and then serum starved overnight. Cells were treated (left panel) with Camptothecin (CPT) for 16 hours. Data shown are from a 20 second exposure using a chemiluminescence imaging system.

Application Notes

Suggested Application

Multiplexed Protein Detection; Detection of Relative Protein Expression; Detecting Patterns of Cytokine Expression; Biomarker/ Key Factor Screening; Identifying Key Factors; Confirming a Biological Process

Kit Components

Mouse Apoptosis Signaling Pathway Antibody Array C1 Membranes
Blocking Buffer
Detection Antibody Cocktail
1,000X HRP-Anti-Rabbit IgG Concentrate
20X Wash Buffer I Concentrate
20X Wash Buffer II Concentrate
2X Cell Lysis Buffer Concentrate
Detection Buffer C
Detection Buffer D
8-Well Incubation Tray w/ Lid
Protease Inhibitor Cocktail
100x Phosphatase Inhibitor Cocktail I
Phosphatase Inhibitor Cocktail II
Plastic Sheets
Array Map Template
Manual

Other Materials Required

Pipettors, pipet tips and other common lab consumables
Orbital shaker or oscillating rocker
Tissue Paper, blotting paper or chromatography paper
Adhesive tape or Saran Wrap
Distilled or de-ionized water
A chemiluminescent blot documentation system (such as UVP's ChemiDoc-It^® or EpiChem II Benchtop Darkroom or GE's ImageQuant^¢ LAS 4000 or Amersham Imagers 600 and 680), X-ray Film and a suitable film processor, or another chemiluminescent detection system.

Protocol Outline

Block membranes
Incubate with Sample
Incubate with Detection Antibody Cocktail
Incubate with HRP-Conjugated anti-IgG
Incubate with Detection Buffers
Image with chemiluminescent imaging system
Perform densitometry and analysis

Storage/Stability

For best results, store the entire kit frozen at -20^°C upon arrival. Stored frozen, the kit will be stable for at least 6 months which is the duration of the product warranty period. Once thawed, store array membranes at -20^°C and all other reagents undiluted at 4^°C for no more than 3 months.

Radulovic, Katarina, et al. "NLRP6 Deficiency in CD4 T Cells Decreases T Cell Survival Associated with Increased Cell Death." The Journal of Immunology 203.2 (2019): 544-556.
Species: Mouse
Sample type: Cell Lysate
Liu F, Qiu H, Xue M, et al. MSC-secreted TGF-? regulates lipopolysaccharide-stimulated macrophage M2-like polarization via the Akt/FoxO1 pathway. Stem Cell Res Ther. 2019;10(1):345. Published 2019 Nov 26. doi:10.1186/s13287-019-1447-y
Species: Mouse
Sample type: Cell Lysate

It is important to select the protein concentration as a positive sample before the experiment. I recommend that western blot or another experiment should be confirmed an expression level before using a kit. Optionally, for a clear image under the low background, I recommend using a high sensitivity ECL solution.

Hyuk-Kwon Kwon
Yale University 06/28/2019, 05:19 PM

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