Membrane-based antibody array detects 19 phosphorylated or cleaved human apoptosis proteins. Suitable for all liquid sample types but intended for use with cell and tissue lysates.
Product Description
Specifications
| Size | 2 Sample Kit, 4 Sample Kit, 8 Sample Kit |
|---|---|
| Species | Human |
| Quantitative/Semi-Quantitative | Semi-Quantitative |
| Number of Targets Detected | 19 |
| Compatible Sample Types | Cell Culture Supernatants, Plasma, Serum, Tissue Lysates, Cell Lysates |
| Solid Support | Membrane |
| Method Of Detection | Chemiluminescence |
| Design Principle | Sandwich-based |
| Research Area | Apoptosis, Akt Signaling, HIF-1 alpha Signaling, IGF Signaling, JAK/STAT Signaling, MAPK Signaling, mTOR Signaling, PI3K-AKT Signaling, TGF-beta Signaling, Wnt/beta-Catenin Signaling, p53 Signaling, NFkB Signaling |
| Estimated Lead Time | 1-2 business days |
| Shipping Type | Blue ice |
| Storage | -20°C |
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| Scroll over each target protein for more information | ||||
|---|---|---|---|---|
|
Akt (P-Ser473)
(PKB (P-Ser473))
|
ATM (P-Ser1981)
|
BAD (P-Ser112)
|
Caspase-3 (Cleaved Asp175)
|
Caspase-7 (Cleaved Asp198)
|
|
CHK1 (P-Ser296)
|
CHK2 (P-Thr68)
|
eIF-2a (P-Ser52)
|
ERK1 (P-T202/Y204)/ERK2 (P-Y185/Y187)
|
HSP27 (P-Ser82)
|
|
IKBa (P-Ser32)
|
JNK (P-Thr183/Tyr185)
|
NFKBP65 (P-Ser536)
|
PARP1 (Cleaved Asp214/Gly215)
|
p27 (P-Thr198)
|
|
p38 (P-Thr180/Tyr182)
|
P53 (P-Ser15)
|
SMAD2 (Ser245/250/255)
|
TAK1 (P-Ser412)
|
|
Typical Results
Figure 1: HeLa cells were grown to 80% confluency and then serum starved overnight. Cells were either untreated (bottom panel) or treated (top panel) with Staurosporine (STS) for 3 hours. Data shown are from a 20 second exposure using a chemiluminescence imaging system.
Figure 2: Jurkat cells were either untreated (bottom panel) or treated (top 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
- Human Apoptosis Signaling 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.
Vergani E, Dugo M, Cossa M, Frigerio S, Di Guardo L, Gallino G, Mattavelli I, Vergani B, Lalli L, Tamborini E, Valeri B, Gargiuli C, Shahaj E, Ferrarini M, Ferrero E, Gomez Lira M, Huber V, Vecchio MD, Sensi M, Leone BE, Santinami M, Rivoltini L, Rodolfo M, Vallacchi V. miR-146a-5p impairs melanoma resistance to kinase inhibitors by targeting COX2 and regulating NFkB-mediated inflammatory mediators. Cell Commun Signal. 2020 Sep 23;18(1):156. doi: 10.1186/s12964-020-00601-1. PMID: 32967672; PMCID: PMC7510138.
Species:
Human
Sample Type:
Cell Lysate
Hii, LW., Lim, SH.E., Leong, CO. et al. The synergism of Clinacanthus nutans Lindau extracts with gemcitabine: downregulation of anti-apoptotic markers in squamous pancreatic ductal adenocarcinoma. BMC Complement Altern Med 19, 257 (2019). https://doi.org/10.1186/s12906-019-2663-9
Species:
Human
Sample Type:
Conditioned Media (cell culture media)
Smolensky D, Rhodes D, McVey DS, et al. High-Polyphenol Sorghum Bran Extract Inhibits Cancer Cell Growth Through ROS Induction, Cell Cycle Arrest, and Apoptosis. J Med Food. 2018;21(10):990-998. doi:10.1089/jmf.2018.0008
Species:
Human
Sample Type:
Cell Lysate (liver heptaocellular carcinoma)
Bao X, Shi J, Xie F, et al. Proteolytic Release of the p75NTR Intracellular Domain by ADAM10 Promotes Metastasis and Resistance to Anoikis. Cancer Res. 2018;78(9):2262-2276. doi:10.1158/0008-5472.CAN-17-2789
Species:
Human
Sample Type:
Cell Lysate (carcinoma cells)
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Great productWas investigating ulcerative colitis serum exomes and drug ADS04 on HPECs and any downstream apoptosis. The kit was easy to use and gave a really good, clean output at the end
from UCLA,
on
Great productThe human apoptosis array provides a fast and comprehensive glance about the key signaling involved in the apoptotic pathway. It's a one shot experiment with certain controls already on the blot, pretty simple to execute and normalize. Would recommend it.from Moffitt Cancer Center,
on
Great productThis apoptosis kit was very user-friendly and gave detailed instructions on how to analyze findings using a software like Excel. This assay was very helpful for examining multiple gene targets simultaneously, so we got a lot of data out of 1 experiment. Negative and positive controls were included on the blots, which was useful in subsequent analyses.Notes: the Caspase3 antibody seemed a lot more sensitive than other antibodies on the blot. Also, some of the gene names in the kit manual differed from those listed on the website, so keep that in mind when analyzing data to ensure that the correct gene is being referred to.from Medical College of Wisconsin,
on
Great productIt seems antibodies are not evenly attached on the membrane. Some signals show up outside of spots. It has high background when I follow up the suggested procedures. I cannot see any benefit of this array compared to other array such as apoptosis array or other company's similar product.from Penn State College of Medicine,
on
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