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Assay Format

Pathway:

Akt Signaling
JAK/STAT Signaling
MAPK Signaling
NFkB Signaling
TGF-beta Signaling

Number of Targets Detected:

Species Detected:

Human

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 | AKT1S1 | AMPK1 | AND | ATF2 | ATM | BAD | CDKN1B | CREB1 | EGFR 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
4E-BP1 (P-Thr36) Gene Symbol: EIF4EBP1 Gene ID: 1978 Accession #: Q13541 ELISA Kit: RayBio Human/Mouse Phospho-4E-BP1 (T36) ELISA	AKT (P-Ser473) Gene Symbol: Akt1 Akt Rac Gene ID: 11651 Accession #: P31750 ELISA Kit: RayBio Human/Mouse/Rat Phospho-Akt (S473) ELISA	AMPKa (P-Thr172) Gene Symbol: AMPK1 Gene ID: 5562 Accession #: P54645 ELISA Kit: RayBio	ATF2 (P-Thr69/71) Gene Symbol: ATF2 CREB2 CREBP1 Gene ID: 1386; Accession #: P15336 ELISA Kit: RayBio Human and Mouse Phospho-ATF2 (Thr69/71) ELISA	ATM (P-Ser1981) Gene Symbol: ATM Gene ID: 472; Accession #: Q13315 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	C-Fos (P-Thr232) Gene Symbol: FOS G0S7 Gene ID: 2353; Accession #: P01100 ELISA Kit: RayBio Human Phospho-C-Fos (Thr232) ELISA	c-Jun (P-Ser73) Gene Symbol: JUN Gene ID: 3725; Accession #: P05412 ELISA Kit: RayBio	CREB (P-Ser133) Gene Symbol: CREB1 Gene ID: 1385 Accession #: P16220 ELISA Kit: RayBio Human Phospho-CREB (S133) ELISA	EGFR (P-Ser1070) Gene Symbol: EGFR ERBB ERBB1 HER1 Gene ID: 1956 Accession #: P00533 ELISA Kit: RayBio Human Phospho-EGFR (S1070) ELISA
eIF-2a (P-Ser51) Gene Symbol: EIF2S1 EIF2A Gene ID: 1965; Accession #: P05198 ELISA Kit: RayBio	ERK1/2 (P-Thr202/P-Tyr204/P-Tyr185/P-Tyr187) Gene Symbol: AND ERK1 ERK2 MAPK1 MAPK3 PRKM1 PRKM2 PRKM3 Gene ID: 5595 5594 Accession #: P27361 P28482 ELISA Kit: RayBio Custom Human Phosphotyrosine ERK1/2 ELISA	GSK-3 alpha (P-Ser21) Gene Symbol: GSK3A Gene ID: 2931 Accession #: P49840 ELISA Kit: RayBio Human/Mouse/Rat Phospho-GSK3a (S21) ELISA	GSK-3 beta (P-Ser9) Gene Symbol: GSK3B Gene ID: 2932; Accession #: P49841 ELISA Kit: RayBio Human Phospho-GSK3b (S9) ELISA	HDAC2 (P-Ser394) Gene Symbol: HDAC2 Gene ID: 3066 Accession #: Q92769 ELISA Kit: RayBio Human, Mouse and Rat Phospho-HDAC2 (S394) ELISA
HDAC4 (P-Ser632) Gene Symbol: HDAC4 KIAA0288 Gene ID: 9759; Accession #: P56524 ELISA Kit: RayBio Human/Mouse/Rat Phospho-HDAC4 (S632)	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	JAK1 (P-Tyr1022) Gene Symbol: JAK1 JAK1A JAK1B Gene ID: 3716 Accession #: P23458 ELISA Kit: RayBio Human/Mouse/Rat Phospho-Jak1 (Y1022) ELISA	JAK2 (P-Tyr1007/1008) Gene Symbol: JAK2 Gene ID: 3717 Accession #: O60674 ELISA Kit: RayBio Human Phospho-JAK2 (Y1007/1008) 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	MEK (P-Ser217/221) Gene Symbol: MAP2K1 MEK1 PRKMK1 Gene ID: 5604 Accession #: Q02750 ELISA Kit: RayBio Human/Mouse/Rat Phospho-Mek (S217/S221) ELISA	MKK3 (P-Ser189) Gene Symbol: MAP2K3 MEK3 MKK3 PRKMK3 SKK2 Gene ID: 5606 Accession #: P46734 ELISA Kit: RayBio Human/Mouse/Rat Phospho-MKK3 (S189) ELISA	MKK6 (P-Ser207) (MEK6) Gene Symbol: MAP2K6 MEK6 MKK6 PRKMK6 SKK3 Gene ID: 5608 Accession #: P52564 ELISA Kit: RayBio Human/Mouse/Rat Phospho-MKK6 (S207) ELISA	MSK1 (P-Ser376) Gene Symbol: RPS6KA5 MSK1 Gene ID: 9252; Accession #: O75582 ELISA Kit: RayBio Human Phospho-MSK1 (S376) ELISA
MSK2 (P-Ser360) Gene Symbol: RPS6KA4 MSK2 Gene ID: 8986 Accession #: O75676 ELISA Kit: RayBio Human Phospho-MSK2 (S360) ELISA	mTOR (P-Ser2448) Gene Symbol: MTOR FRAP FRAP1 FRAP2 RAFT1 RAPT1 Gene ID: 2475 Accession #: P42345 ELISA Kit: RayBio Human Phospho-mTOR (S2448) ELISA	NF-kB (P-Ser536) Gene Symbol: RELA NFKB3 Gene ID: 5970 Accession #: Q04206 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	P70S6K (P-Thr421/Ser424) Gene Symbol: RPS6KB1 STK14A Gene ID: 6198 Accession #: P23443 ELISA Kit: RayBio Human/Mouse/Rat Phospho-P70S6K (T421/S424) ELISA	PDK1 (P-Ser241) Gene Symbol: PDPK1 PDK1 Gene ID: 5170 Accession #: O15530 ELISA Kit: RayBio Human, Mouse and Rat Phospho-PDK1 (Ser241) ELISA	PRAS40 (P-Thr246) Gene Symbol: AKT1S1 PRAS40 Gene ID: 84335 Accession #: Q96B36 ELISA Kit: RayBio Human Phospho-PRAS40 (T246) ELISA	PTEN (P-Ser380) Gene Symbol: PTEN MMAC1 TEP1 Gene ID: 5728 Accession #: P60484 ELISA Kit: RayBio Human/Mouse/Rat Phospho-PTEN (S380) ELISA
Raf-1 (P-Ser301) Gene Symbol: RAF1 RAF Gene ID: 5894 Accession #: P04049 ELISA Kit: RayBio Human/Mouse/Rat Phospho-Raf-1 (Ser301) ELISA	RPS6 (P-Ser235/236) Gene Symbol: RPS6 OK/SW-cl.2 Gene ID: 6194 Accession #: P62753 ELISA Kit: RayBio Human/Mouse/Rat Phospho-RPS6 (S235/S236) ELISA	RSK1 (P-Ser380) Gene Symbol: RPS6KA1 MAPKAPK1A RSK1 Gene ID: 6195 Accession #: Q15418 ELISA Kit: RayBio Human Phospho-RSK1 (S380) ELISA	RSK2 (P-Ser386) Gene Symbol: RPS6KA3 ISPK1 MAPKAPK1B RSK2 Gene ID: 6197 Accession #: P51812 ELISA Kit: RayBio Human, Mouse and Rat Phospho-RSK2 (Ser386) ELISA	SHP-1 (P-Ser591) Gene Symbol: PTPN6 HCP PTP1C Gene ID: 5777 Accession #: P29350 ELISA Kit: RayBio Human and Mouse Phospho-SHP1 (Ser591) ELISA
SHP2 (P-Tyr542) Gene Symbol: PTPN11 PTP2C SHPTP2 Gene ID: 5781 Accession #: Q06124 ELISA Kit: RayBio Human/Mouse/Rat Phospho-SHP2 (Y542) ELISA	SMAD1 (P-Ser463/465) Gene Symbol: SMAD1 BSP1 MADH1 MADR1 Gene ID: 4086; Accession #: Q15797 ELISA Kit: RayBio Human Phospho-SMAD1 (SER463/465)	SMAD2 (P-Ser245/250/255) Gene Symbol: SMAD2 MADH2 MADR2 Gene ID: 4087; Accession #: Q15796 ELISA Kit: RayBio Human/Mouse/Rat Phospho-SMAD2 (SER245/250/255) ELISA	SMAD4 (P-Thr277) Gene Symbol: SMAD4 DPC4 MADH4 Gene ID: 4089; Accession #: Q13485 ELISA Kit: RayBio Human Phospho-SMAD4 (Thr277) ELISA	SMAD5 (P-Ser463/465) Gene Symbol: SMAD5 MADH5 Gene ID: 4090; Accession #: Q99717 ELISA Kit: RayBio Human Phospho-SMAD5 (SER463/465) ELISA
Src (P-Tyr419) Gene Symbol: SRC SRC1 Gene ID: 6714 Accession #: P12931 ELISA Kit: RayBio Human Phospho-SRC (Y419) ELISA	STAT1 (P-Ser727) Gene Symbol: STAT1 Gene ID: 6772 Accession #: P42224 ELISA Kit: RayBio Human/Mouse Phospho-Stat 1 (S727) ELISA	STAT2 (P-Tyr689) Gene Symbol: STAT2 Gene ID: 6773 Accession #: P52630 ELISA Kit: RayBio Human Phospho-STAT2 (Y689) ELISA	STAT3 (P-Tyr705) Gene Symbol: STAT3 APRF Gene ID: 6774 Accession #: P40763 ELISA Kit: RayBio Human/Mouse/Rat Phospho-Stat 3 (Y705) ELISA	STAT5 (P-Tyr694) Gene Symbol: STAT5A STAT5 Gene ID: 6776 Accession #: P42229 ELISA Kit: RayBio Human Phosphotyrosine STAT5 ELISA
STAT6 (P-Tyr641) Gene Symbol: STAT6 Gene ID: 6778 Accession #: P42226 ELISA Kit: RayBio Human Phospho-STAT6 (Y641) ELISA	TAK1 (P-Ser412) Gene Symbol: MAP3K7 TAK1 Gene ID: 6885; Accession #: O43318 ELISA Kit: RayBio Human Phospho-TAK1 (Ser412) ELISA	TBK1 (P-Ser172) Gene Symbol: TBK1 NAK Gene ID: 29110; Accession #: Q9UHD2 ELISA Kit: RayBio Human, Mouse and Rat Phospho-TBK1 (S172) ELISA	TYK2 (P-Tyr1054) Gene Symbol: TYK2 Gene ID: 7297 Accession #: P29597 ELISA Kit: RayBio Human and Mouse Phospho-TYK2 (Tyr1054) ELISA	ZAP70 (P-Tyr292) Gene Symbol: ZAP70 SRK Gene ID: 7535; Accession #: P43403 ELISA Kit: RayBio Human Phosphotyrosine ZAP70 ELISA

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 Phosphorylation Pathway Profiling Array C55 Membranes
Blocking Buffer
Detection Antibody Cocktail
500X 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 and 1X Blocking Buffer at -20^°C and all other reagents undiluted at 4^°C for no more than 3 months.

Kang J, Chun J, Hwang JS, Pan C, Li J, Boese AC, Young I, Malin CM, Kang Y, Gibbons DL, Sica G, Fu H, Ramalingam SS, Jin L, Kang S. EGFR-phosphorylated GDH1 harmonizes with RSK2 to drive CREB activation and tumor metastasis in EGFR-activated lung cancer. Cell Rep. 2022 Dec 13;41(11):111827. doi: 10.1016/j.celrep.2022.111827. PMID: 36516759; PMCID: PMC9813823.
Species: Human
Sample type: Cell Lysate
Zhang, Dan, et al. "MALAT1 is involved in the pathophysiological process of PCOS by modulating TGF? signaling in granulosa cells." Molecular and cellular endocrinology 499 (2020): 110589.
Species: Human
Sample type: Cell Lysate
Zhang D, Tang HY, Tan L, Zhao DM. MALAT1 is involved in the pathophysiological process of PCOS by modulating TGF? signaling in granulosa cells. Mol Cell Endocrinol. 2020 Jan 1;499:110589. doi: 10.1016/j.mce.2019.110589. Epub 2019 Sep 23. PMID: 31557499.
Species: Human
Sample type: Cell Lysate

It was easy to test multiple phosphorylation targets with limited sample quantity and workflow was really easy and uncomplicated. I used primary human endothelium cells with 100 ug of protein for each array and signals were good with 10-sec exposure.

Jay Mishra
University of Wisconsin-Madison 07/05/2021, 07:56 PM

This kit works pretty well for my samples. I used 700 ug of my samples (cell lysates) and performed exactly according to the instructional protocol and I got a very strong signal. The only reason that I didn't give it 5 stars was we had to purchase their software to analyze the results. As we already spent nearly $1000, we should be able to use the software for free, at least one time free.

Jianhui Ma
Kalya Pharma 07/15/2019, 11:43 AM

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