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Human Follistatin Quantikine ELISA Kit

R&D Systems | Catalog # DFN00

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Key Product Details

Assay Length

5.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (10 µL), Serum (100 µL), EDTA Plasma (100 µL), Follicular Fluid (10 µL)

Sensitivity

83 pg/mL

Assay Range

250-16000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Follicular Fluid)
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Product Summary for Human Follistatin Quantikine ELISA Kit

The Quantikine Human Follistatin Immunoassay is a 5.5 hour solid-phase ELISA designed to measure human Follistatin (FS288, FS300, and FS315) in follicular fluid, cell culture supernates, serum, and plasma. It contains Sf 21-expressed recombinant human Follistatin and antibodies raised against the recombinant factor. It has been shown to accurately quantitate recombinant human Follistatin. Results obtained using natural Follistatin showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural Follistatin.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

Specificity

Natural and recombinant human Follistatin

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

Interference observed with 1 or more available related molecules.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

Cell Culture Supernates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 1850 5363 10734 1645 4850 10256
Standard Deviation 59 136 181 139 312 584
CV% 3.2 2.5 1.7 8.4 6.4 5.7

EDTA Plasma, Follicular Fluid, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 1996 6047 11999 1888 5604 11626
Standard Deviation 53 144 239 173 399 918
CV% 2.7 2.4 2.0 9.2 7.1 7.9

Recovery for Human Follistatin Quantikine ELISA Kit

The recovery of Follistatin spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 97 91-107
EDTA Plasma (n=5) 96 87-102
Serum (n=5) 95 87-104

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Follistatin were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human Follistatin ELISA Linearity

Scientific Data Images for Human Follistatin Quantikine ELISA Kit

Human Follistatin ELISA Cell Culture Supernate Standard Curve

Human Follistatin ELISA Cell Culture Supernate Standard Curve

Human Follistatin ELISA Follicular Fluid/Serum/Plasma Standard Curve

Human Follistatin ELISA Follicular Fluid/Serum/Plasma Standard Curve

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: Follistatin

Follistatin (FST) is a secreted glycoprotein that binds and neutralizes several TGF-beta superfamily proteins including Activin, BMP-6, BMP-7, and GDF-8/Myostatin. It also binds Angiogenin and regulates hematopoietic stem cell adhesion to Fibronectin. Some Follistatin binding partners will also bind Follistatin-like proteins such as FSL-3. Of three follistatin isoforms, the full-length mature follistatin (FST315) is the most abundant and the sole form in plasma, but has lower binding affinity for both activins and heparins than alternative forms. The acidic tail is missing in the splice variant FST288, which shows highest affinity, and partial in the proteolytically produced FST303, which shows intermediate affinity. Genetic deletion of follistatin in mice, or expression of only the FST288 form, is lethal perinatally due to defects of lung, skin and musculoskeletal system. Expression of only the FST315 isoform allows survival, with defects in vascularization and female fertility.

Alternate Names

FS, FST

Entrez Gene IDs

10468 (Human); 14313 (Mouse)

Gene Symbol

FST

Additional Follistatin Products

Product Documents for Human Follistatin Quantikine ELISA Kit

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.

Product Specific Notices for Human Follistatin Quantikine ELISA Kit

For research use only

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Citations for Human Follistatin Quantikine ELISA Kit

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    Sample Tested: Serum
    Verified Customer | Posted 04/07/2020
    Human Follistatin Quantikine ELISA Kit DFN00
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    Sample Tested: Cell Culture Supernates
    Verified Customer | Posted 08/06/2018
  • Human Follistatin Quantikine ELISA Kit
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    Sample Tested: Cell Culture Supernates
    Verified Customer | Posted 11/07/2016
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Protocols

View specific protocols for Human Follistatin Quantikine ELISA Kit (DFN00):

Refer to the product for complete assay procedure.

The conjugate must remain at 2-8 °C until use. Bring all other reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
    3. 100 µL Assay Diluent
  3.   Add 100 µL of Assay Diluent to each well.
    4. 100 µL Standard, Control, or Sample
  4.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at 2-8 °C for 3 hours.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL cold Conjugate
  6.   Add 200 µL of cold Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 2 hours.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well.
  9.   For Cell Culture Supernate Samples: Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
    For Serum, Plasma, & Follicular Fluid Samples: Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    10. 50 µL Stop Solution
  10. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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