Charged multivesicular body protein 2b (CHMP2.5) (Chromatin-modifying protein 2b) (CHMP2b) (Vacuolar protein sorting-associated protein 2-2) (Vps2-2) (hVps2-2)
Accession Number
Q9UQN3
Gene Symbols
CHMP2B|CGI-84
Gene Id
25978
Quantitative/Semi-Quantitative
Quantitative
Compatible Sample Types
Cell Culture Supernatants, Plasma, Serum
Solid Support
96-well Microplate **The PCR plate used is a 0.2mL, non-skirted 96-well plate (ThermoFisher, cat. no.: AB0600)
Method Of Detection
qPCR
Design Principle
Sandwich-based
Sensitivity
Typically 10x lower than traditional ELISA
Risk-Free Guarantee
We offer a 100% guarantee on all ELISA Kits or C-Series Membrane Array.
NEED HELP RUNNING THIS ASSAY?Full testing service for only $5 More per plateREQUEST QUOTE | MORE INFO
Introduction
The RayBio®Immuno Quantitative Enzyme Linked ImumunoSorbent Assay (IQELISA™) is an innovative new 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 RayBio® Custom Human CHMP2B IQELISA™ Kit is a modified ELISA assay with high sensitivity qPCR readout for the quantitative measurement of Human CHMP2B in serum, plasma, and cell culture supernatants. This assay employs an antibody specific for Human CHMP2B coated on a 96-well PCR plate. Standards and samples are pipetted into the wells and CHMP2B 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.
Typical Data
Application Notes
Kit Components
CHMP2B Microplate (Item A)**: 96 well PCR plate coated with anti-Human CHMP2B
Wash Buffer I Concentrate (20x) (Item B): 25 ml of 20x concentrated solution
Standards (Item C): 2 vials of recombinant Human CHMP2B
Assay Diluent B (Item E): 15 ml of 5x concentrated buffer
Detection Affinity Reagent for CHMP2B (Item F): 2 vials of a 4x concentrated solution of anti-Human CHMP2B 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).
Other Materials Required
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
Protocol Outline
Prepare all reagents, samples and standards as instructed
Add 25 µl standard or sample to each well. Incubate for 1.5 - 2.5 hours at room temperature
Add 25 µl detection antibody to each well. Incubate 1 hour at room temperature
Add 50 µl of IQELISA™ Detection Reagent to each well. Incubate 1 hour
Add 10 µl Primer solution and 10 µl of PCR master mix to each well
Run real-time PCR
Storage/Stability
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°C. Note: the kit can be used within one year if the whole kit is stored at -20°C. Avoid repeated freeze-thaw cycles.
