Human TGF-alpha Quantikine ELISA Kit

R&D Systems | Catalog # DTGA00

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL), Human Milk (50 µL)

Sensitivity

7.1 pg/mL

Assay Range

15.6-1000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Human Milk)
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Product Summary for Human TGF-alpha Quantikine ELISA Kit

The Quantikine Human TGF-alpha Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human TGF-alpha in cell culture supernates, serum, plasma, and human milk. It contains E. coli-expressed recombinant human TGF-alpha and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human TGF-alpha 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 naturally occurring human TGF-alpha.

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 TGF-alpha

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. Cross-species reactivity not tested.

Interference

No significant interference observed with 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, EDTA Plasma, Heparin Plasma, Human Milk, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 91.7 215 419 93.1 221 415
Standard Deviation 3.8 9.6 11.1 6.3 10.7 18.1
CV% 4.1 4.5 2.6 6.8 4.8 4.4

Recovery for Human TGF-alpha Quantikine ELISA Kit

The recovery of TGF-alpha spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 97 92-108
EDTA Plasma (n=4) 99 88-114
Heparin Plasma (n=4) 103 88-114
Serum (n=4) 103 96-114

Linearity

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

Human TGF-alpha ELISA Linearity

Scientific Data Images for Human TGF-alpha Quantikine ELISA Kit

Human TGF-alpha ELISA Standard Curve

Human TGF-alpha ELISA 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: TGF-alpha

TGF-alpha is a member of the EGF family of cytokines that are synthesized as transmembrane precursors with soluble forms released by proteolytic cleavage. Membrane-bound proTGF-alpha is biologically active and plays a role in mediation of cell-cell adhesion and in juxtacrine stimulation of adjacent cells. Expression of TGF-alpha is widespread in tumors and transformed cells. TGF-alpha is also expressed in normal tissues during embryogenesis and in adult tissues, including pituitary, brain, keratinocytes and macrophages. TGF-alpha binds and activates the EGF receptor. It induces fibroblast proliferation, epithelial development, keratinocyte migration, and angiogenesis.

Long Name

Transforming Growth Factor alpha

Alternate Names

TGFA, TGFalpha

Entrez Gene IDs

7039 (Human)

Gene Symbol

TGFA

Additional TGF-alpha Products

Product Documents for Human TGF-alpha 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 TGF-alpha 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 TGF-alpha Quantikine ELISA Kit

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  • Human TGF-alpha Quantikine ELISA Kit
    Name: Rahul Mallick
    Sample Tested: HUVEC human umbilical vein endothelial cells
    Verified Customer | Posted 08/26/2020
  • Human TGF-alpha Quantikine ELISA Kit
    Name: Leslie Priddy
    Sample Tested: Serum
    Verified Customer | Posted 04/01/2018

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Protocols

View specific protocols for Human TGF-alpha Quantikine ELISA Kit (DTGA00):

Refer to the product for complete assay procedure.

Bring all 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. 50 µL Standard, Control, or Sample
  4.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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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