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Verwendungszweck
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Mouse Immunoquantitative (PCR-Based) Kremen-1 ELISA Kit for cell culture supernatants, plasma, and serum samples.
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Proben
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Cell Culture Supernatant, Plasma, Serum
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Analytische Methode
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Semi-Quantitative
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Nachweismethode
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qPCR
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Produktmerkmale
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IQ-ELISAs employ specific capture antibodies coated on a 96-well PCR plate. Standards and samples are pipetted into the wells, the target protein in the standards and samples binds to the immobilized antibody. The wells are washed and the detection affinity reagent is added to the wells where it binds to any captured antigen. The wells are washed, and primers and PCR master mix are added to each well. The plate is placed into a real time PCR instrument for cycling and measurement of DNA amplification. The cycle number where amplification is detected is proportional to the amount of affinity detection reagent that bound to captured antigen in each well.
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Bestandteile
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- Kremen-1 Microplate (Item A): 96 well PCR plate coated with anti-Mouse Kremen-1
- Wash Buffer I Concentrate (20x) (Item B): 25 ml of 20x concentrated solution
- Standards (Item C): 2 vials of recombinant Mouse Kremen-1
- Assay Diluent A (Item D): 30 ml diluent buffer, 0.09% sodium azide as preservative. For Standard/Sample (serum/plasma) diluent
- Assay Diluent B (Item E): 15 ml of 5x concentrated buffer. For Standard/Sample (cell culture medium/urine) diluent
- Detection Affinity Reagent for Kremen-1 (Item F): 2 vials of a 4x concentrated solution of anti-Mouse Kremen-1 affinity reagent
- IQELISA Detection Reagent (Item G): 1.4ml of a 10x concentrated stock
- Primer Solution (Item I): 1.7 ml vial
- PCR Master Mix (Item J): 1.2 ml vial
- PCR Preparation buffer (Item K): 1ml vial of 10x concentrated buffer
- Final Wash Buffer (Item L): 10ml vial of 10x concentrated buffer
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Benötigtes Material
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- 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
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Plasmids, Primers & others
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KREMEN1 products on genomics-online (e.g. as negative or positive controls)
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