c-MET ELISA Kit

Catalog No. ABIN1672789

Mouse c-MET ELISA Kit Colorimetric assay for quantification of Mouse c-MET.

Quick Overview for c-MET ELISA Kit (ABIN1672789)

Target: c-MET (MET) (Met Proto-Oncogene (MET))

Binding Specificity: AA 25-306, AA 307-929

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 62.5-4000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA)

Product Details c-MET ELISA Kit

Minimum Detection Limit: 62.5 pg/mL

Purpose: For quantitative detection of mouse C-MET in cell culture supernates, serum and plasma(heparin, EDTA).

Brand: PicoKine™

Analytical Method: Quantitative

Specificity: Natural and recombinant mouse C-MET

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <5pg/mL

Components:

• 96-well plate precoated with anti- mouse C-MET antibod - 1
• Lyophilized recombinant mouse C-MET standar - 10ng/tubex2
• Biotinylated anti- mouse C-MET antibod - 130μl(dilution 1:100)
• Avidin-Biotin-Peroxidase Complex (ABC - 130μl(dilution 1:100)
• Sample diluent buffe - 30 ml
• Antibody diluent buffe - 12ml
• ABC diluent buffe - 12ml
• TMB color developing agen - 10ml
• TMB stop solutio - 10ml

Material not included: Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl

Immunogen: Expression system for standard: sf21
Immunogen sequence: E25-R306(alpha)&S307-N929(beta)

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Plate: Pre-coated

Protocol: mouse C-MET ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for C-MET has been precoated onto 96-well plates. Standards(sf21, E25-R306(α) &S307-N929(β)) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for C-MET is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the mouse C-MET amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 4000pg/mL, 2000pg/mL,1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL mouse C-MET standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of mouse cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each mouse C-MET standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(ng/ml): 0.51, Standard deviation: 0.022, CV(%): 4.3
• Sample 2: n=16, Mean(ng/ml): 1.7, Standard deviation: 0.092, CV(%): 5.4
• Sample 3: n=16, Mean(ng/ml): 2.8, Standard deviation: 0.146, CV(%): 5.2,
• Sample 1: n=24, Mean(ng/ml): 0.57, Standard deviation: 0.029, CV(%): 5.1
• Sample 2: n=24, Mean(ng/ml): 1.8, Standard deviation: 0.137, CV(%): 7.6
• Sample 3: n=24, Mean(ng/ml): 3.2, Standard deviation: 0.221, CV(%): 6.9

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

Target Details for c-MET

Target: c-MET (MET) (Met Proto-Oncogene (MET))

Alternative Name: MET

Background: C-Met(MET or MNNG HOS Transforming gene) is a proto-oncogene that encodes a protein known as hepatocyte growth factor receptor(HGFR). MET proto-oncogene has a total length of 125,982 bp, and it is located in the 7q31 locus of chromosome 7. MET is a membrane receptor that is essential for embryonic development and wound healing. Activation of MET triggers mitogenesis, and morphogenesis.

Gene ID: 17295

UniProt: A4FUV6

Pathways: RTK Signaling, Carbohydrate Homeostasis, Synaptic Membrane, Signaling of Hepatocyte Growth Factor Receptor