Mouse L-Selectin IQELISA™ Kit

RayBio® Mouse Immunoquantitative (PCR-Based) L-Selectin ELISA Kit for cell culture supernatants, plasma, and serum samples. 20-25 business day lead time

As low as $547.00
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Code:
IQM-LSelectin
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Product Description

Specifications

Size 1 Plate Kit, 2 Plate Kit, 5 Plate Kit
Species Mouse
Protein Name / Synonyms L-selectin
Accession Number Q3TCF3
Gene Symbols SELL
Gene Id 20343
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 3.5 pg/ml (Anticipated minimum sensitivity. Assay may detect lower levels of antigen.)

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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® Mouse L-Selectin IQELISA™ Kit is a modified ELISA assay with high sensitivity qPCR readout for the quantitative measurement of Mouse L-Selectin in serum, plasma, and cell culture supernatants. This assay employs an antibody specific for Mouse L-Selectin coated on a 96-well PCR plate. Standards and samples are pipetted into the wells and L-Selectin 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
  • L-Selectin Microplate (Item A)**: 96 well PCR plate coated with anti-Mouse L-Selectin
  • Wash Buffer I Concentrate (20x) (Item B): 25 ml of 20x concentrated solution
  • Standards (Item C): 2 vials of recombinant Mouse L-Selectin
  • Assay Diluent B (Item E): 15 ml of 5x concentrated buffer
  • Detection Affinity Reagent for L-Selectin (Item F): 2 vials of a 4x concentrated solution of anti-Mouse L-Selectin 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
  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

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.
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