c-MET IQ-ELISA Kit

Catalog No. ABIN5680031

This qPCR ELISA kit is designed for the quantitative measurement of Human c-MET.

Quick Overview for c-MET IQ-ELISA Kit (ABIN5680031)

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

Reactivity: Human

Detection Method: qPCR

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Culture Supernatant, Plasma, Serum

Product Details c-MET IQ-ELISA Kit

Purpose: Human Immunoquantitative (PCR-Based) HGFR ELISA Kit for cell culture supernatants, plasma, and serum samples.

Analytical Method: Semi-Quantitative

Sensitivity: 6 pg/mL

Characteristics: The Immuno Quantitative Enzyme Linked ImumunoSorbent Assay (IQELISA) is an innovative assay that combines the specificity and ease of use of an ELISA with the sensitivity of real-time PCR. This results in an assay that is simultaneously familiar and cutting edge and enables the use of lower sample volumes while also providing more sensitivity. The Human HGFR IQELISA kit is a modified ELISA assay with high sensitivity qPCR readout for the quantitative measurement of human HGFR in serum, plasma, and cell culture supernatants. This assay employs an antibody specific for human HGFR coated on a 96-well PCR plate. Standards and samples are pipetted into the wells and HGFR present in a sample is bound to the wells by the immobilized antibody. The wells are washed and a detection affinity molecule is added to the plates. After washing away unbound detection affinity molecule, primers and a PCR master mix are added to the wells and data is collected using qPCR. Ct values obtained from the qPCR are then used to calculate the amount of antigen contained in each sample, where lower Ct values indicate a higher concentration of antigen.

Components:

• HGFR Microplate (Item A): 96 well PCR plate coated with anti-human HGFR
• Wash Buffer I Concentrate (20x) (Item B): 20x concentrated solution
• Standards (Item C): recombinant human HGFR
• Assay Diluent A (Item D): diluent buffer, 0.09% sodium azide as preservative. For Standard/Sample (serum/plasma) diluent
• Assay Diluent B (Item E): 5x concentrated buffer. For Standard/Sample (cell culture medium/urine) diluent
• Detection Affinity Reagent for HGFR (Item F): 4x concentrated solution of anti-human HGFR affinity reagent
• Primer Solution (Item G)
• PCR Master Mix (Item H)
• PCR Preparation buffer (Item I): 10x concentrated buffer
• Final Wash Buffer (Item J): 10x concentrated buffer

Material not included:

• Real-time PCR instrument, Bio-Rad recommended
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• 100 mL and 1 liter graduated cylinders
• Absorbent paper
• Distilled or deionized water
• Log-log graph paper or computer and software for data analysis
• Tubes to prepare standard or sample dilutions
• Heating block or water bath capable of 80 °C

Application Details

Application Notes: The Immuno-Quantitative ELISA (IQELISA) kits are an innovative assay platform that combines the specificity and ease of use of an ELISA with the sensitivity of real-time PCR. This results in an assay that is simultaneously familiar and cutting edge and enables the use of only 1/10th the sample volume while also providing 10x more sensitivity than a traditional ELISA.

Comment:

Features:
- Excellent for follow up/confirmation of antibody array data
- Pre-coated 96 well plate
- Strip plate format minimizes waste
- Complete kit includes all necessary reagents
- 10x more sensitive than ELISA
- Reproducible and reliable
- Optimized for serum, plasma, urine and cell culture medium
- 1/10th the sample volume of an ELISA

Research Applications:
- Detection of quantitative protein levels in biological fluids
- Validation of antibody array results
- Validation of biomarker discovery studies

Plate: Pre-coated

Protocol: 1. Prepare all reagents, samples and standards as instructed
2. Add 25 μL standard or sample to each well. Incubate for 2.5 hours at room temperature or overnight at 4 °C
3. Add 25 μL detection affinity reagent to each well. Incubate 1 hour at room temperature
4. Add 15μL Primer solution to each well
5. Add 10μL of PCR Master Mix to each well
6. Run real-time PCR

Sample Preparation:

Recommended Dilution (serum & plasma): 5 - 100 fold

Assay Precision: Serum spike tests show recovery is 92.5% with a range of 89.7 to 98.1%

Restrictions: For Research Use only

Handling

Storage: 4 °C,-20 °C,-80 °C

Storage Comment: May be stored for up to 6 months at 2° to 8°C from the date of shipment. Standard (recombinant protein) should be stored at -20°C or -80°C (recommended at -80°C) after reconstitution. Opened PCR plate or reagents may be stored for up to 1 month at 2° to 8

Expiry Date: 6 months

Target Details

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

Alternative Name: HGFR

Background: Synonyms: Hepatocyte growth factor receptor (HGF receptor) (EC 2.7.10.1) (HGF/SF receptor) (Proto-oncogene c-Met) (Scatter factor receptor) (SF receptor) (Tyrosine-protein kinase Met), MET

Gene ID: 4233

UniProt: P08581

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