Human Gas6 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY885B
Ancillary Products Available
Human Gas6 ELISA Standard Curve
1 Image
Product Details
Procedure
Citations (4)
FAQs
Supplemental Products
Reviews (2)

Human Gas6 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
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 Growth Arrest-Specific Protein 6 (Gas6). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, plasma, and urine samples. Diluents for complex matrices, such as serum, plasma, and urine, 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)

Data Example

Human Gas6 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: Gas6

Gas6 (Growth Arrest Specific 6) is a secreted protein that contains an extensively gamma-carboxylated N-terminal Gla domain and is dependent upon vitamin K for activity. Gas6 signals through the receptors Axl, Dtk/Tyro3, and Mer and is inhibited by shed soluble forms of Axl and Mer. Gas6 protects cells from stress-induced apoptosis and promotes phagocytosis of apoptotic cell debris, promotes platelet activation but inhibits inflammatory cytokine production, and inhibits VEGF-induced angiogenesis, and regulates tumor cell invasiveness.

Long Name:
Growth-arrest-specific Protein 6
Entrez Gene IDs:
2621 (Human); 14456 (Mouse)
Alternate Names:
AXLLG; AXLLGAXL stimulatory factor; AXSFAXL receptor tyrosine kinase ligand; DKFZp666G247; FLJ34709; Gas6; GAS-6; growth arrest-specific 6; growth arrest-specific protein 6

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 Gas6 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.

4 Citations: Showing 1 - 4
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  1. Axl is required for TGF-?2-induced dormancy of prostate cancer cells in the bone marrow
    Sci Rep, 2016;6(0):36520.
    Species: Human
    Sample Types: Cell Lysates
  2. Role of Growth arrest-specific gene 6-Mer axis in multiple myeloma.
    Authors: Waizenegger J, Ben-Batalla I, Weinhold N, Meissner T, Wroblewski M, Janning M, Riecken K, Binder M, Atanackovic D, Taipaleenmaeki H, Schewe D, Sawall S, Gensch V, Cubas-Cordova M, Seckinger A, Fiedler W, Hesse E, Kroger N, Fehse B, Hose D, Klein B, Raab M, Pantel K, Bokemeyer C, Loges S
    Leukemia, 2015;29(3):696-704.
    Species: Human
    Sample Types: Bone Marrow Plasma
  3. Plasma concentrations predict aortic expression of growth-arrest-specific protein 6 in patients undergoing coronary artery bypass grafting.
    Authors: Lee, Chien-Hs, Shieh, Yi-Shing, Tsai, Chien-Su, Hung, Yi-Jen, Tsai, Yi-Ting, Lin, Chih-Yua
    PLoS ONE, 2013;8(11):e79452.
    Species: Human
    Sample Types: Plasma
  4. GAS6/Mer axis regulates the homing and survival of the E2A/PBX1-positive B-cell precursor acute lymphoblastic leukemia in the bone marrow niche.
    Authors: Shiozawa Y, Pedersen EA, Taichman RS
    Exp. Hematol., 2010;38(2):132-40.
    Species: Human
    Sample Types: Cell Culture Supernates

FAQs

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Reviews for Human Gas6 DuoSet ELISA

Average Rating: 4.5 (Based on 2 Reviews)

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Human Gas6 DuoSet ELISA
By Anonymous on 09/16/2017
Sample Tested: PC-3 human prostate cancer cell line

Human Gas6 DuoSet ELISA
By Anonymous on 06/08/2017
Sample Tested: Cell Culture Media