Human Integrin alpha 4 beta 1 DuoSet ELISA

R&D Systems | Catalog # DY5668-05

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Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

125-8000 pg/mL

Sample Type

Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet

Reactivity

Human

Human Integrin alpha 4 beta 1 DuoSet ELISA 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
View all Integrin alpha 4 beta 1 ELISA Kits »

Product Summary for Human Integrin alpha 4 beta 1 DuoSet ELISA

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Integrin α4β1. 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 Specifications

Assay Format

96-well strip plate (sold separately)

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Specificity

Please see the product datasheet

Label

HRP

Scientific Data Images for Human Integrin alpha 4 beta 1 DuoSet ELISA

Human Integrin alpha a4 beta 1 Standard Curve

Kit Contents for Human Integrin alpha 4 beta 1 DuoSet ELISA

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

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008C) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Reagent Diluent*

Blocking Buffer*

Substrate Solution: ELISA TMB Substrate (Catalog # DY999B or DY999B-250)

Stop Solution: Methanesulfonic acid (Catalog # DY994B or DY994B-250)

Microplates: (Catalog # DY990), or equivalent

Plate Sealers: (Catalog # DY992), or equivalent

*For the recommended Reagent Diluent and Blocking Buffer for a specific DuoSet ELISA Development Kit, refer to the product datasheet.

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: Integrin alpha 4 beta 1

Integrin alpha 4 beta 1, also known as VLA-4, is an Integrin family adhesion receptor that shares the beta 1 subunit with eleven other family members and the alpha 4 subunit with Integrin alpha 4 beta 7. The noncovalent heterodimer of alpha 4/CD49d and beta 1/CD29 subunits mediates cell adhesion to VCAM-1/CD106 on other cells and the CS-1 fragment of Fibronectin in the extracellular matrix. Leukocytes (except for neutrophils), erythroid precursors, and some non-hematopoietic cells such as epicardial, endothelial and smooth muscle precursors, Schwann cells, and chorionic cells express a4b1. In the adult, a4b1 primarily regulates immune cell migration. Circulating leukocyte a4b1 is rapidly activated by inflamed endothelial cells that present VCAM-1 and chemokines such as CXCL12/SDF-1. This activation facilitates rolling, firm adhesion, and extravasation. During immune cell activation, a4b1 can function as a costimulatory molecule.

 

Alternate Names

CD49d, IA4

Entrez Gene IDs

3676 (Human)

Gene Symbol

ITGA4

Additional Integrin alpha 4 beta 1 Products

Product Documents for Human Integrin alpha 4 beta 1 DuoSet ELISA

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human Integrin alpha 4 beta 1 DuoSet ELISA

For research use only

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Protocols

View specific protocols for Human Integrin alpha 4 beta 1 DuoSet ELISA (DY5668-05):

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.

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