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Human FGF basic/FGF2/bFGF Quantikine ELISA Kit Best Seller

R&D Systems, part of Bio-Techne | Catalog # DFB50

R&D Systems, part of Bio-Techne
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DFB50
PDFB50
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (100 µL), Serum (100 µL), EDTA Plasma (100 µL)

Sensitivity

3 pg/mL

Assay Range

10-640 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)

Product Summary for Human FGF basic/FGF2/bFGF Quantikine ELISA Kit

The Quantikine Human FGF basic Immunoassay kit is a 4.5 hour solid phase ELISA designed to measure human FGF basic in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human FGF basic and antibodies raised against the recombinant factor. It has been shown to quantitate recombinant human FGF basic accurately. Results obtained using natural human FGF basic 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 human FGF basic.

Product Specifications

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Human

Specificity

Natural and recombinant human FGF basic.  This assay cross-reacts 40% with natural Bovine FGF-basic, 0.15% with natural Bovine FGF-acidic, 17% with recombinant Bovine FGF-basic, 18% with recombinant Mouse FGF-basic and 57% with recombinant Rat FGF-basic.

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. Cross-species reactivity observed with 1 or more species tested.

Interference

No significant interference observed with available related molecules.

Sample Values

Serum/Plasma - Samples from apparently healthy volunteers were evaluated for the presence of human FGF basic in this assay. No medical histories were available for the donors used in this study.

Sample TypeMean of Detectable (pg/mL)% DetectableRange (pg/mL)
SerumND0ND
EDTA plasma13.510ND-14.6
ND=Non-detectable

Cell Culture Supernates:
Human peripheral blood mononuclear cells (5 x 106 cells/mL) were cultured in RPMI supplemented with 5% fetal bovine serum, 50 μM beta-mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells were stimulated with 10 μg/mL PHA. Aliquots of the culture supernates were removed on days 1 and 5 and assayed for levels of natural human FGF basic. All samples measured below the lowest standard, 10 pg/mL.

K562 human chronic myelogenous leukemia cells (4 x 104 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. An aliquot of the cell culture supernate was removed on day 3, assayed for natural human FGF basic, and measured 34 pg/mL.

PC-3 human prostate cancer cells were cultured in MEM supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells were stimulated with 10 μg TNF-alpha. An aliquot of the cell culture supernate was removed, assayed for natural human FGF basic, and measured 27 pg/mL.

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, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 28.8 104 234 32.8 108 229
Standard Deviation 2.8 3.2 7.1 3.0 8.0 17.3
CV% 9.7 3.1 3.0 9.1 7.4 7.6

Recovery for Human FGF basic/FGF2/bFGF Quantikine ELISA Kit

The recovery of FGF basic spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 108 94-119
EDTA Plasma (n=5) 103 90-124
Serum (n=5) 106 86-124

Linearity

To assess the linearity of the assay, the following samples spiked with high concentrations of FGF basic were diluted with Calibrator Diluent and then assayed.

Human FGF basic ELISA Linearity

Scientific Data Images for Human FGF basic/FGF2/bFGF Quantikine ELISA Kit

Human FGF basic ELISA Standard Curve

Human FGF basic 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: FGF basic/FGF2/bFGF

FGF basic/FGF2/bFGF is a growth factor that functions in angiogenesis, wound healing, tissue repair, learning and memory, and the morphogenesis of heart, bone, and brain. It is upregulated in response to inflammatory stimuli and in many tumors. FGF basic/FGF2/bFGF binds to FGFR1c and 2c. Its bioactivity is modulated by a number of other binding partners including heparin, Integrin alpha V beta 3, soluble FGFR1, FGF-BP, free gangliosides, Thrombospondin, Pentraxin 3/TSG-14, Fibrinogen, alpha 2-Macroglobulin, PDGF, and CXCL4/PF4. These molecules act as cellular coreceptors or adhesion partners, extracellular matrix decoys or reservoirs, and soluble scavengers or chaperones. In particular, the interaction of FGF basic/FGF2/bFGF with cell surface heparan sulfate proteoglycans (HSPG) is required for the binding and activation of FGF receptors.

Long Name

Fibroblast Growth Factor basic

Alternate Names

bFGF, FGF-2, FGF2, HBGF-2, Prostatropin

Entrez Gene IDs

2247 (Human); 14173 (Mouse); 281161 (Bovine); 403857 (Canine); 100033955 (Equine)

Gene Symbol

FGF2

Additional FGF basic/FGF2/bFGF Products

Product Documents for Human FGF basic/FGF2/bFGF Quantikine ELISA Kit

Certificate of Analysis

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

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human FGF basic/FGF2/bFGF Quantikine ELISA Kit

For research use only

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⚠ 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.