Custom Human NRG1 Isoform SMDF IQELISA™ Kit

• NRG1 Isoform SMDF Microplate (Item A)**: 96 well PCR plate coated with anti-Human NRG1 Isoform SMDF
• Wash Buffer I Concentrate (20x) (Item B): 25 ml of 20x concentrated solution
• Standards (Item C): 2 vials of recombinant Human NRG1 Isoform SMDF
• Assay Diluent B (Item E): 15 ml of 5x concentrated buffer
• Detection Affinity Reagent for NRG1 Isoform SMDF (Item F): 2 vials of a 4x concentrated solution of anti-Human NRG1 Isoform SMDF affinity reagent
• IQELISA™ Detection Reagent (Item G): 1 ml of a 10x concentrated stock
• Primer Solution (Item I): 1.5 ml vial
• PCR Master Mix (Item J): 1.4 ml vial
• PCR Preparation buffer (Item K): 1 ml vial of 10x concentrated buffer
• Final Wash Buffer (Item L): 10 ml vial of 10x concentrated buffer

**The PCR plate used is a 0.2 ml, non-skirted 96-well plate (ThermoFisher, cat. no.: AB0600). Please ensure compatibility with your PCR machine prior to purchase. For additional information contact technical support (techsupport@raybiotech.com).

• 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

1. Prepare all reagents, samples and standards as instructed
2. Add 25 µl standard or sample to each well. Incubate for 1.5 - 2.5 hours at room temperature
3. Add 25 µl detection antibody to each well. Incubate 1 hour at room temperature
4. Add 50 µl of IQELISA™ Detection Reagent to each well. Incubate 1 hour
5. Add 10 µl Primer solution and 10 µl of PCR master mix to each well
6. Run real-time PCR