Human PDGF R beta DuoSet ELISA

Catalog # Availability Size / Price Qty
DY385-05
Ancillary Products Available
Human PDGF-R beta  ELISA Standard Curve
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Product Details
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Human PDGF R beta DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Range
125.0 - 8,000 pg/mL
Sufficient Materials
For five 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Platelet-Derived Growth Factor Receptor beta (PDGF-Rβ). The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Scientific Data

Human PDGF-R beta ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: PDGF R beta

PDGF R beta is a transmembrane receptor tyrosine kinase (RTK) that primarily binds to the PDGF-BB isoform of the growth factor. Ligand binding induces receptor homodimerization or heterodimerization with PDGF R alpha followed by autophosphorylation and signal transduction. PDGF receptor signaling is attenuated by SHP-2 phosphatase activity and by receptor endocytosis and lysosomal degradation. The expression of the alpha and beta receptors is independently regulated in various cell types. A soluble splice variant of PDGF R beta binds PDGF with high affinity and functions as a potent PDGF antagonist.

Long Name:
Platelet-derived Growth Factor Receptor beta
Entrez Gene IDs:
5159 (Human); 18596 (Mouse); 24629 (Rat)
Alternate Names:
beta-type platelet-derived growth factor receptor; CD140 antigen-like family member B; CD140b antigen; CD140b; EC 2.7.10; EC 2.7.10.1; JTK12; PDGF R beta; PDGFR; PDGFR1; PDGFRB; PDGF-R-beta; platelet-derived growth factor receptor, beta polypeptide

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

FAQs

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