Human BDNF IQELISA™ Kit

$522.00
IQH-BDNF-1

RayBio® Human Immunoquantitative (PCR-Based) BDNF ELISA Kit for cell culture supernatants, plasma, and serum samples. 10-15 business day lead time.

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Antigen Information

Accession Number:
  • P23560
Gene ID:
  • 627
Gene Symbol:
  • BDNF
Protein Name & Synonyms:
Brain-derived neurotrophic factor (BDNF) (Abrineurin)
Target Species:
  • Human

Assay Format

Sensitivity:
8 pg/ml (Anticipated minimum sensitivity. Assay may detect lower levels of antigen.)
Number of Targets Detected:
1
Species Detected:
  • Human
Compatible Sample Types:
  • Cell Culture Supernatants
  • Plasma
  • Serum
Design Principle:
  • Sandwich-based
Method of Detection:
qPCR
Quantitative/Semi-Quantitative:
Quantitative
Solid Support:
96-well Microplate

Product Specifications

Size:
1, 2, or 5 x 96-Well Strip Microplate Kit

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

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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Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.