Human/Mouse MAPK Phosphorylation Array

RayBio® C-Series Human/Mouse MAPK Phosphorylation Antibody Array 1 Kit. Detects 17 phosphorylated proteins in human or mouse samples. Suitable for all liquid sample types but intended for use with cell and tissue lysates.

$342.26  
AAH-MAPK-1-2
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Assay Format

Pathway:
  • Akt Signaling
  • HER/ErbB Signaling
  • IGF Signaling
  • JAK/STAT Signaling
  • MAPK Signaling
  • NFkB Signaling
  • PI3K-AKT Signaling
  • Wnt/beta-Catenin Signaling
Number of Targets Detected:
17
Species Detected:
  • Human
  • 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:
AKT1 | CREB1 | GSK3A | GSK3B | HSPB1 | MAP2K1 | MAP2K3 | MAP2K6 | MAPK1 | MAPK14   View more

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)
(PKB (P-Ser473))
CREB (P-Ser133)
ERK1 (P-T202/Y204)/ERK2 (P-Y185/Y187)
GSK3a (P-Ser21)
GSK3b (P-Ser9)
HSP27 (P-Ser82)
JNK (P-Thr183)
MEK (P-Ser217/Ser221)
MKK3 (P-Ser189)
MKK6 (P-Ser207)
MSK2 (P-Ser360)
mTOR (P-Ser2448)
p38 (P-Thr180/Tyr182)
P53 (P-Ser15)
P70S6K (P-Thr421/Ser424)
RSK1 (P-Ser380)
RSK2 (P-Ser386)
  • Gene Symbol: RPS6KA3 ISPK1 MAPKAPK1B RSK2
  • Gene ID: 10432
  • Accession #: P51812
     

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/Mouse MAPK Phosphorylation 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
  1. Block membranes
  2. Incubate with Sample
  3. Incubate with Detection Antibody Cocktail
  4. Incubate with HRP-Conjugated anti-IgG
  5. Incubate with Detection Buffers
  6. Image with chemiluminescent imaging system
  7. Perform densitometry and analysis

Typical Data

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 250 nM PMA for 20 minutes. Data shown are from a 20 second exposure using a chemiluminescence imaging system.
Figure 2 - HepG2 cells were grown to 80% confluency and then serum starved overnight. Cells were either untreated (bottom panel) or treated (top panel) with 25 ng/mL of recombinant human IL-1β for 30 minutes. Data shown are from a 20 second exposure using a chemiluminescence imaging system.

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 and 1X Blocking Buffer at -20°C and all other reagents undiluted at 4°C for no more than 3 months.
  1. Poling, Holly M., et al. "Mechanically induced development and maturation of human intestinal organoids in vivo." Nature biomedical engineering 2.6 (2018): 429-442.
    Species: Human
    Sample type: Tissue Lysate
  2. Xie Z, Peng M, Lu R, Meng X, Liang W, Li Z, Qiu M, Zhang B, Nie G, Xie N, Zhang H, Prasad PN. Black phosphorus-based photothermal therapy with aCD47-mediated immune checkpoint blockade for enhanced cancer immunotherapy. Light Sci Appl. 2020 Sep 15;9:161. doi: 10.1038/s41377-020-00388-3. PMID: 33014356; PMCID: PMC7492464.
    Species: Mouse
    Sample type: Tissue Lysate
  3. Yu S., Zhong Q., Chen R., et al. CRLF1 promotes malignant phenotypes of papillary thyroid carcinoma by activating the MAPK/ERK and PI3K/AKT pathways. Cell Death Dis. 2018 Mar 7;9(3):371. doi: 10.1038/s41419-018-0352-0.
    Species: Human
    Sample type: Cell Lysate (Human thyroid cell lines, normal and cancerous; IHH-4, PTC, etc)
  4. Yu, Shi-Tong, et al. "CRLF1 promotes malignant phenotypes of papillary thyroid carcinoma by activating the MAPK/ERK and PI3K/AKT pathways." Cell death & disease 9.3 (2018): 1-13.
    Species: Human
    Sample type: Cell Lysate
  5. Schuett J., Schuett H., Oberoi R., et al. NADPH oxidase NOX2 mediates TLR2/6-dependent release of GM-CSF from endothelial cells. FASEB J. 2017 Mar 8. pii: fj.201600729R. doi: 10.1096/fj.201600729R
    Species: Human
    Sample type: Cell Lysate (Human umbilical cord endothelial cells)
  6. Schuett, Jutta, et al. "NADPH oxidase NOX2 mediates TLR2/6?dependent release of GM?CSF from endothelial cells." The FASEB Journal 31.6 (2017): 2612-2624.
    Species: Human
    Sample type: Cell Lysate
  7. Yang Z, Zou W, Pan Y, Yong KT, Li L, Wang X, Liu D, Chen T, Xue D, Lin G. PEGylated CuInS2/ZnS quantum dots inhibit neurite outgrowth by downregulating the NGF/p75NTR/MAPK pathway. Ecotoxicol Environ Saf. 2021 Jan 1;207:111378. doi: 10.1016/j.ecoenv.2020.111378. Epub 2020 Oct 3. PMID: 33022524.
    Species: Rat
    Sample type: Cell Lysate
  8. Dhani, Shanel, et al. "Fusaric acid alters Akt and ampk signalling in c57bl/6 mice brain tissue." Food and Chemical Toxicology (2020): 111252.
    Species: Mouse
    Sample type: Tissue Lysate (brain tissue)
  9. Sarsour, Ehab H., et al. "Arachidonate 12-lipoxygenase and 12-hydroxyeicosatetraenoic acid contribute to stromal aging-induced progression of pancreatic cancer." Journal of Biological Chemistry 295.20 (2020): 6946-6957.
    Species: Human
    Sample type: Cell Lysate (pancreatic cells)
  10. L'Huillier C, Jarbeau M, Achamrah N, et al. Glutamine, but not Branched-Chain Amino Acids, Restores Intestinal Barrier Function during Activity-Based Anorexia. Nutrients. 2019;11(6):1348. Published 2019 Jun 15. doi:10.3390/nu11061348
    Species: Mouse
    Sample type: Tissue Lysate (Colon tissue )
1.Experimental conditions and sample type
a. In vitro: Human breast cancer cell line (hBCC)
b. In vivo: hBCC grew up in nude mouse bone + mouse bone (human/mouse protein mixed)

Pros: pAKT1, pCREB, pGSK3b,p70S6K, pRSK1,2, pMEK1, pp38, and pMKK3,6 are good. The experimental process is very easy. Easy to reproduce the results

Cons: pERK1/2 is a signal relatively weak compared to the Cell signaling antibody.

Yale University
This assay is very straightforward and provides good results for brain samples.

Emory
These arrays are what I was looking for. They are easy to use, very clean membranes with super sharp results. These arrays are a great second step for large scale analysis such RNAseq.

The University of Iowa
Easy to use and convenient way to look at multiple targets.

Midwestern University
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Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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