The kit is designed for in vitro quantitative measurement of Mouse CD27 in Cell Culture Supernatant, Plasma, Serum.
Description
Mouse Immunoquantitative (PCR-Based) CD27 antigen ELISA Kit for cell culture supernatants, plasma, and serum samples.
Applications
ELISA
Application Notes
The Immuno-Quantitative ELISA 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
The Immuno-Quantitative ELISA kits are an innovative assay platform that combines the specificity and ease of use of an ELISA with the sensitivity of real-time PCR. Also called immuno-PCR, this detection platform results in an assay that is simultaneously familiar and cutting edge. Compared to traditional ELISA, quantitative ELISA enables the use of only 1/10th the sample volume while also providing 10x more sensitivity.
Target
CD27
Reactivity
Mouse
Detection Method
qPCR
Method Type
Sandwich ELISA
Analytical Method
Semi-Quantitative
Sample Type
Cell Culture Supernatant, Plasma, Serum
Components
CD27 antigen Microplate (Item A): 96 well PCR plate coated with anti-Mouse CD27 antigen. Wash Buffer I Concentrate (20x) (Item B): 25 ml of 20x concentrated solution. Standards (Item C): 2 vials of recombinant Mouse CD27 antigen. Assay Diluent A (Item D): 30 ml diluent buffer, 0.09% sodium azide as preservative. Assay Diluent B (Item E): 15 ml of 5x concentrated buffer. Detection Affinity Reagent for CD27 antigen (Item F): 2 vials of a 4x concentrated solution of anti-Mouse CD27 antigen affinity reagent. Quantitative ELISA 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.
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.
Sample Volume
25 µL
Plate
Pre-coated
Storage
4 °C,-20 °C,-80 °C
Storage Comment
May be stored for up to 6 months at 2°C 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°C to 8°C.
Expiry Date
6 months
Note
May be stored for up to 6 months at 2°C 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°C to 8°C.
Restrictions
For Research Use only
Alternative Name
Tumor Necrosis Factor Receptor Superfamily, Member 7 (TNFRSF7)
Production of Molecular Mediator of Immune Response, Cancer Immune Checkpoints
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 25μL of quantitative ELISA Detection Reagent to each well. Incubate 1 hour. 5.Add 15μL Primer solution 10μL of PCR master mix to each well. 6.Run real-time PCR.