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Manual / SDS

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Assay Format

Pathway:

Akt Signaling
HIF-1 alpha 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:

4 or 8 Sample Kit

Protein Information

Gene Symbols:

MMP1
MMP10
MMP13
MMP2
MMP3
MMP8
MMP9
TIMP1
TIMP2
TIMP4

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
MMP-1 Gene Symbol: MMP1 Gene ID: 4312 Accession #: P03956 ELISA Kit: RayBio Human MMP-1 ELISA	MMP-2 Gene Symbol: MMP2 Gene ID: 4313 Accession #: P08253 ELISA Kit: RayBio Human MMP-2 ELISA	MMP-3 Gene Symbol: MMP3 Gene ID: 4314 Accession #: P08254 ELISA Kit: RayBio Human MMP-3 ELISA	MMP-8 Gene Symbol: MMP8 Gene ID: 4317 Accession #: P22894 ELISA Kit: RayBio Human MMP-8 ELISA	MMP-9 Gene Symbol: MMP9 Gene ID: 4318 Accession #: P14780 ELISA Kit: RayBio Human MMP-9 ELISA
MMP-10 Gene Symbol: MMP10 Gene ID: 4319 Accession #: P09238 ELISA Kit: RayBio Human MMP-10 ELISA	MMP-13 Gene Symbol: MMP13 Gene ID: 4322 Accession #: P45452 ELISA Kit: RayBio Human MMP-13 ELISA	TIMP-1 Gene Symbol: TIMP1 Gene ID: 7076 Accession #: P01033 ELISA Kit: RayBio Human TIMP-1 ELISA	TIMP-2 Gene Symbol: TIMP2 Gene ID: 7077 Accession #: P16035 ELISA Kit: RayBio Human TIMP-2 ELISA	TIMP-4 Gene Symbol: TIMP4 Gene ID: 7079 Accession #: Q99727 ELISA Kit: RayBio Human TIMP-4 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 Matrix Metalloproteinase Antibody Antibody Array C1 Membranes
Blocking Buffer
Wash Buffer 1
Wash Buffer 2
Biotinylated Detection Antibody Cocktail
Streptavidin-Conjugated HRP
Detection Buffer C
Detection Buffer D
Lysis Buffer
8-Well Incubation Tray
Plastic Sheets
Array Templates
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 Biotinylated Detection Antibody Cocktail
Incubate with HRP-Conjugated Streptavidin
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.

Cheng YC, Chiang HY, Cheng SJ, Chang HW, Li YJ, Shieh SY. Loss of the tumor suppressor BTG3 drives a pro-angiogenic tumor microenvironment through HIF-1 activation. Cell Death Dis. 2020 Dec 11;11(12):1046. doi: 10.1038/s41419-020-03248-5. PMID: 33311481; PMCID: PMC7732837.
Species: Mouse
Sample type: Conditioned media
Tinaburri L, Valente C, Teson M, Minafò YA, Cordisco S, Guerra L, Dellambra E. The Secretome of Aged Fibroblasts Promotes EMT-Like Phenotype in Primary Keratinocytes from Elderly Donors through BDNF-TrkB Axis. J Invest Dermatol. 2020 Sep 12:S0022-202X(20)32062-5. doi: 10.1016/j.jid.2020.08.019. Epub ahead of print. PMID: 32931807.
Species: Human
Sample type: Conditioned Media
Kc P, Shah M, Shaik R, Hong Y, Zhang G. Preseeding of Mesenchymal Stem Cells Increases Integration of an iPSC-Derived CM Sheet into a Cardiac Matrix. ACS Biomater Sci Eng. 2020 Dec 14;6(12):6808-6818. doi: 10.1021/acsbiomaterials.0c00788. Epub 2020 Nov 4. PMID: 33320624.
Species: Human
Sample type: Conditioned Media
Chien YC., Liu LC., Ye HY., et al. EZH2 promotes migration and invasion of triple-negative breast cancer cells via regulating TIMP2-MMP-2/-9 pathway. Am J Cancer Res. 2018; 8(3): 422434.PMCID: PMC5883093
Species: Human
Sample type: Conditioned Media
Chien YC, Liu LC, Ye HY, Wu JY, Yu YL. EZH2 promotes migration and invasion of triple-negative breast cancer cells via regulating TIMP2-MMP-2/-9 pathway. Am J Cancer Res. 2018 Mar 1;8(3):422-434. PMID: 29636998; PMCID: PMC5883093.
Species: Human
Sample type: Conditioned media
Perrigue P., Silva M., Warden C., et al. The histone demethylase jumonji coordinates cellular senescence including secretion of neural stem cell-attracting cytokines. Mol Cancer Res. 2015 Apr;13(4):636-50. doi: 10.1158/1541-7786.MCR-13-0268
Species: Human
Sample type: Conditioned Media
Salhi A., Farhadian J., Giles K., et al. RSK1 Activation Promotes Invasion in Nodular Melanoma. Am J Pathol. 2015 Mar;185(3):704-16. doi: 10.1016/j.ajpath.2014.11.021.
Species: Human
Sample type: Tissue Lysate
Huang J., Yang B., Xiang T., et al. Diallyl disulfide inhibits growth and metastatic potential of human triple-negative breast cancer cells through inactivation of the ?-catenin signaling pathway. Mol Nutr Food Res. 2015 Jun;59(6):1063-75. doi: 10.1002/mnfr.201400668.
Species: Human
Sample type: Conditioned Media
Carduner L., Leroy-Dudal J., Picot CR., Gallet O., Carreiras F., et al. Ascites-induced shift along epithelial-mesenchymal spectrum in ovarian cancer cells: enhancement of their invasive behavior partly dependant on ?v integrins. Clinical and Experimental Metastasis. Jun 2014 Epub Ahead of Print.
Species: Human
Sample type: Peritoneal Fluid
Ichikawa T., Sugiura H., Koarai A., Minakata Y., Kikuchi T., Morishita Y., et al. TLR3 Activation Augments Matrix Metalloproteinase Production through Reactive Nitrogen Species Generation in Human Lung Fibroblasts. The Journal of Immunology April 23, 2014 1302919 [Epub Ahead of Print]
Species: Human
Sample type: Conditioned Media

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2 8 well assays were used for an experiment and it worked well. I liked having the instructions available via web however, I used an old printed out manual that I received previously.

Niki
Indiana University School of Dentistry 04/04/2021, 06:38 PM

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