Mouse VCAM-1/CD106 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY643
Ancillary Products Available
VCAM-1/CD106 ELISA Kit ELISA
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Product Details
Procedure
Citations (5)
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Mouse VCAM-1/CD106 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4 hours 40 minutes (after plate preparation)
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 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 mouse VCAM-1. 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)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

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 0.05% Tween® 20 in PBS, pH 7.2-7.4

Block Buffer: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Reagent Diluent: 0.1% BSA, 0.05% Tween 20 in Tris-buffered Saline (20 mM Trizma base, 150 mM NaCI) pH 7.2-7.4, 0.2 μm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

 

Data Example

Mouse VCAM-1 / CD106 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: VCAM-1/CD106

VCAM-1 (Vascular Cell Adhesion Molecule-1; CD106) is a transmembrane molecule that mediates the adhesion of immune cells to the vascular endothelium during inflammation. It binds to several integrins including alpha4/beta1 (VLA-4), alpha4/beta7, alpha9/beta1, and alphaD/beta2. It is expressed constitutively in the bone marrow where it regulates T cell, B cell, and hematopoietic progenitor cell migration. A soluble form VCAM-1 can promote monocyte chemotaxis.

Long Name:
Vascular Cell Adhesion Molecule 1
Entrez Gene IDs:
7412 (Human); 22329 (Mouse)
Alternate Names:
CD106 antigen; CD106; DKFZp779G2333; INCAM-100; L1CAM; MGC99561; vascular cell adhesion molecule 1; vascular cell adhesion protein 1; V-CAM 1; VCAM1; VCAM-1

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 of Block Buffer 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.

Citations for Mouse VCAM-1/CD106 DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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  1. Combination of a high-fat diet with sweetened condensed milk exacerbates inflammation and insulin resistance induced by each separately in mice
    Authors: LN Masi, AR Martins, AR Crisma, CL do Amaral, MR Davanso, TDA Serdan, RDC da Cunha d, MM Cruz, MIC Alonso-Val, RP Torres, J Mancini-Fi, JNB Pereira, MM da Silva R, EA Liberti, SM Hirabara, R Curi
    Sci Rep, 2017;7(1):3937.
    Species: Mouse
    Sample Types: Tissue Homogenate
  2. Imbalance of the Vanin-1 Pathway in Systemic Sclerosis
    J Immunol, 2016;0(0):.
    Species: Mouse
    Sample Types: Serum
  3. Neurokinin-1 receptor antagonist treatment protects mice against lung injury in polymicrobial sepsis.
    Authors: Hegde A, Zhang H, Moochhala SM, Bhatia M
    J. Leukoc. Biol., 2007;82(3):678-85.
    Species: Mouse
    Sample Types: Plasma
  4. Excreted urinary mediators in an animal model of experimental immune nephritis with potential pathogenic significance.
    Authors: Wu T, Xie C, Bhaskarabhatla M, Yan M, Leone A, Chen SS, Zhou XJ, Putterman C, Mohan C
    Arthritis Rheum., 2007;56(3):949-59.
    Species: Mouse
    Sample Types: Urine
  5. Elevated urinary VCAM-1, P-selectin, soluble TNF receptor-1, and CXC chemokine ligand 16 in multiple murine lupus strains and human lupus nephritis.
    Authors: Wu T, Xie C, Wang HW, Zhou XJ, Schwartz N, Calixto S, Mackay M, Aranow C, Putterman C, Mohan C
    J. Immunol., 2007;179(10):7166-75.
    Species: Human
    Sample Types: Urine

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