Human Activin A DuoSet ELISA

R&D Systems | Catalog # DY338

R&D Systems
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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 Activin A 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 Activin A ELISA Kits »

Product Summary for Human Activin A DuoSet ELISA

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Activin A. 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)

Sample Volume Required

100 µL

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Specificity

Please see the product datasheet

Label

HRP

Scientific Data Images for Human Activin A DuoSet ELISA

Human Activin A ELISA Standard Curve

Human Activin A ELISA Standard Curve

Kit Contents for Human Activin A 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

1M Urea in PBS, pH 5.0-5.8

*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: Activin A

Activin A is an important member of the TGF-beta superfamily that is produced by many different cell types and is essential for proper development. Bioactive Activin A exists as a disulfide-linked homodimer that recognizes and binds to four Activin receptors, Activin RIA and RIB, and Activin RIIA and RIIB. This ligand-receptor binding event triggers activation of SMAD signaling pathways ultimately regulating a wide variety of functions including stem cell maintenance, self-renewal, and differentiation, B-cell differentiation and proliferation, wound healing, metabolism, and gonadal development.

Activin A is first produced as precursor protein containing a propeptide that gets proteolytically cleaved off leaving the mature A chain. Two A chains assemble by disulfide bond formation creating the bioactive homodimer. The bioavailability of Activin A is regulated by cell-associated decoy molecules (BAMBI, Betaglycan, Cripto), secreted proteins (a2-Macroglobulin, Follistatin, FLRG), and intracellular incorporation of the beta A subunit into Activin AC or Inhibin A. Experimentally, recombinant Activin A is commonly used to maintain proliferative potential of human induced pluripotent stem cells and to promote differentiation of embryonic stem cells into endoderm and pancreatic B cells.

Alternate Names

activin AB alpha polypeptide, Activin beta-A chain, erythroid differentiation factor, Erythroid differentiation protein, follicle-stimulating hormone-releasing protein, FSH-releasing protein, inhibin beta A chain, inhibin beta A subunit, Inhibin, beta-1

Entrez Gene IDs

3624 (Human); 16323 (Mouse); 29200 (Rat)

Gene Symbol

INHBA

Additional Activin A Products

Product Documents for Human Activin A DuoSet ELISA

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human Activin A DuoSet ELISA

For research use only

Citations for Human Activin A DuoSet ELISA

Customer Reviews for Human Activin A DuoSet ELISA (5)

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  • Human Activin A DuoSet ELISA
    Name: Anonymous
    Sample Tested: Human cell conditioned medium
    Verified Customer | Posted 10/01/2024
    Performed ideally the very first time. We used undiluted samples of medium conditioned for 48h. Carefully read the protocol - this one calls for 1M Urea. Color reaction developed in exactly 20min. Our samples measured within the standard curve range suggested (8000-250pg/ml).
    Human Activin A DuoSet ELISA DY338
  • Human Activin A DuoSet ELISA
    Name: APK Inmunometabolismo
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 11/06/2020
    Human Activin A DuoSet ELISA DY338
  • Human Activin A DuoSet ELISA
    Name: Tracy Sherwood
    Sample Tested: horse serum
    Verified Customer | Posted 07/31/2020
    Further testing would be required for cross-reactivity with horse serum
    Human Activin A DuoSet ELISA DY338
  • Human Activin A DuoSet ELISA
    Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 07/08/2020
    Human Activin A DuoSet ELISA DY338
  • Human Activin A DuoSet ELISA
    Name: Anonymous
    Sample Tested: Skeletal muscle
    Verified Customer | Posted 09/30/2016

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Protocols

View specific protocols for Human Activin A DuoSet ELISA (DY338):

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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