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Human ALCAM DuoSet ELISA

Catalog #: DY656
12 Citations Datasheet / COA / SDS

Discontinued Product

DY656 has been discontinued.
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Human ALCAM / CD166 ELISA Standard Curve
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Citations (12)
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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human ALCAM DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
62.5 - 4,000 pg/mL
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 human ALCAM. 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

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, 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)

Scientific Data

N/A Human ALCAM / CD166 ELISA Standard Curve View Larger

Human ALCAM / CD166 ELISA Standard Curve

Product Datasheets

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Product Datasheet
COA
SDS

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: ALCAM/CD166

Activated leukocyte cell adhesion molecule (ALCAM) is a typeI membrane glycoprotein that is a member of the immunoglobulin superfamily. It is also known as CD166, MEMD, SC-1/DM-GRASP/BEN in the chicken, and KG-CAM in the rat.

Long Name:
Activated Leukocyte Cell Adhesion Molecule
Entrez Gene IDs:
214 (Human); 11658 (Mouse); 101867493 (Cynomolgus Monkey)
Alternate Names:
activated leucocyte cell adhesion molecule; activated leukocyte cell adhesion moleculeFLJ38514; ALCAM; CD166 antigen; CD166; MEMDMGC71733

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.

Citations for Human ALCAM 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.

12 Citations: Showing 1 - 10
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  1. Urinary activated leukocyte cell adhesion molecule as a novel biomarker of lupus nephritis histology
    Authors: H Ding, C Lin, J Cai, Q Guo, M Dai, C Mohan, N Shen
    Arthritis Res. Ther., 2020-05-27;22(1):122.
    Species: Human
    Sample Types: Urine
  2. Epigallocatechin-3-Gallate Prevents Acute Gout by Suppressing NLRP3 Inflammasome Activation and Mitochondrial DNA Synthesis
    Authors: HE Lee, G Yang, YB Park, HC Kang, YY Cho, HS Lee, JY Lee
    Molecules, 2019-06-06;24(11):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  3. The plasticity of primary microglia and their multifaceted effects on endogenous neural stem cells in vitro and in vivo
    Authors: SU Vay, LJ Flitsch, M Rabenstein, R Rogall, S Blaschke, J Kleinhaus, N Reinert, A Bach, GR Fink, M Schroeter, MA Rueger
    J Neuroinflammation, 2018-08-13;15(1):226.
    Species: Rat
    Sample Types: Cell Culture Supernates
  4. Neutrophil Migration and Adhesion Molecule Expression after Acute High-Intensity Street Dance Exercise
    Authors: L Borges, A Dermargos, S Gray, MB Barros Sil, V Santos, TC Pithon-Cur, R Gorjão, E Hatanaka
    J Immunol Res, 2018-07-08;2018(0):1684013.
    Species: Human
    Sample Types: Plasma
  5. Activated leukocyte cell adhesion molecule (ALCAM/CD166) regulates T cell responses in a murine model of food allergy
    Authors: YS Kim, MN Kim, KE Lee, JY Hong, MS Oh, SY Kim, KW Kim, MH Sohn
    Clin. Exp. Immunol., 2018-02-20;0(0):.
    Species: Human
    Sample Types: Serum
  6. Monocyte/macrophage and T cell activation markers are not independently associated with MI risk in healthy individuals - results from the HUNT Study
    Authors: T Ueland, LE Laugsand, LJ Vatten, I Janszky, C Platou, AE Michelsen, JK Damås, P Aukrust, BO Åsvold
    Int. J. Cardiol., 2017-06-12;0(0):.
    Species: Human
    Sample Types: Serum
  7. Activated leukocyte cell adhesion molecule regulates the interaction between pancreatic cancer cells and stellate cells
    Mol Med Rep, 2016-08-26;14(4):3627-33.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. JAM-A and ALCAM are therapeutic targets to inhibit diapedesis across the BBB of CD14+CD16+ monocytes in HIV-infected individuals.
    Authors: Williams D, Anastos K, Morgello S, Berman J
    J Leukoc Biol, 2014-11-24;97(2):401-12.
    Species: Human
    Sample Types: Plasma
  9. Activated leukocyte cell adhesion molecule soluble form: a potential biomarker of epithelial ovarian cancer is increased in type II tumors.
    Authors: Carbotti G, Orengo A, Mezzanzanica D, Bagnoli M, Brizzolara A, Emionite L, Puppo A, Centurioni M, Bruzzone M, Marroni P, Rossello A, Canevari S, Ferrini S, Fabbi M
    Int J Cancer, 2012-12-13;132(11):2597-605.
    Species: Human
    Sample Types: Serum
  10. Activated leukocyte cell adhesion molecule expression and shedding in thyroid tumors.
    Authors: Micciche F, Da Riva L, Fabbi M, Pilotti S, Mondellini P, Ferrini S, Canevari S, Pierotti MA, Bongarzone I
    PLoS ONE, 2011-02-22;6(2):e17141.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Integrated proteomic analysis of human cancer cells and plasma from tumor bearing mice for ovarian cancer biomarker discovery.
    Authors: Pitteri SJ, JeBailey L, Faca VM, Thorpe JD, Silva MA, Ireton RC, Horton MB, Wang H, Pruitt LC, Zhang Q, Cheng KH, Urban N, Hanash SM, Dinulescu DM
    PLoS ONE, 2009-11-19;4(11):e7916.
    Species: Human
    Sample Types: Plasma
  12. The ALCAM shedding by the metalloprotease ADAM17/TACE is involved in motility of ovarian carcinoma cells.
    Authors: Rosso O, Piazza T, Bongarzone I
    Mol. Cancer Res., 2007-12-01;5(12):1246-53.
    Species: Human
    Sample Types: Cell Culture Supernates

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