PDGF-AB Heterodimer ELISA Kit

Catalog No. ABIN411345

Product Details

Target: PDGF-AB Heterodimer

Binding Specificity: AA 82-190, AA 87-211

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 31.2-2000 pg/mL

Minimum Detection Limit: 31.2 pg/mL

Application: ELISA

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human PDGF-AB

Brand: PicoKine™

Sample Type: Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA)

Analytical Method: Quantitative

Specificity: Expression system for standard: E.coli
Immunogen sequence: (S87-T211)+(S82-T190)

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <2pg/mL

Material not included: Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl

Immunogen: Expression system for standard: E.coli
Immunogen sequence: (S87-T211)+(S82-T190)

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Comment:

Sequence similarities: Belongs to the PDGF/VEGF growth factor family.

Tissue Specificity: Expressed at high levels in the heart, brain (sustantia nigra), placenta and fetal kidney. Expressed at moderate levels in the brain (hippocampus), skeletal muscle, kidney and lung. .

Plate: Pre-coated

Protocol: human PDGF-AB ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for PDGF-AB has been precoated onto 96-well plates. Standards(E.coli, (S87-T211)+(S82-T190)) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for PDGF-AB is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human PDGF-AB amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 2000pg/mL,1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL human PDGF-AB standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of human cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each human PDGF-AB standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 237, Standard deviation: 11.6, CV(%): 4.9
• Sample 2: n=16, Mean(pg/ml): 532, Standard deviation: 27.7, CV(%): 5.2
• Sample 3: n=16, Mean(pg/ml): 974, Standard deviation: 62.4, CV(%): 6.3,
• Sample 1: n=24, Mean(pg/ml): 254, Standard deviation: 13.97, CV(%): 5.5
• Sample 2: n=24, Mean(pg/ml): 683, Standard deviation: 45.8, CV(%): 6.7
• Sample 3: n=24, Mean(pg/ml): 1205, Standard deviation: 95.2, CV(%): 7.9

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

References

Takeuchi, Kamei, Shinomiya, Sunagawa, Suzuki, Kamoda, Ohtori, Ochi: "Human platelet-rich plasma promotes axon growth in brain-spinal cord coculture." in: Neuroreport, Vol. 23, Issue 12, pp. 712-6, (2012) (PubMed).

Target Details

Target: PDGF-AB Heterodimer

Alternative Name: Platelet-derived growth factor (PDGF) (PDGF-AB)

Synonyms: PDGF-A ELISA Kit, PDGF1 ELISA Kit, platelet derived growth factor subunit A ELISA Kit, PDGFA ELISA Kit

Background:

Protein Function: Growth factor that plays an essential role in the regulation of embryonic development, cell proliferation, cell migration, survival and chemotaxis. Potent mitogen for cells of mesenchymal origin. Required for normal proliferation and recruitment of pericytes and vascular smooth muscle cells in the central nervous system, skin, lung, heart and placenta. Required for normal blood vessel development, and for normal development of kidney glomeruli. Plays an important role in wound healing. Signaling is modulated by the formation of heterodimers with PDGFA (By similarity). .

Background: The platelet-derived growth factor(PDGF) is a mitogen derived from human platelets consisting of two related polypeptides termed A and B chains. The genes for PDGF A chain, B chain/c-sis, and the PDGF receptor are expressed in human malignant glioma cell lines. Normal human endothelial cells in culture express the B chain of PDGF, and that endothelial-derived PDGF B chain is synthesized as a predicted precursor polypeptide of Mr 27,281. The entire B chain of PDGF is highly(96 % ) homologous to a portion of p28sis, the transforming protein of simian sarcoma virus(SSV). It has been suggested that p28sis exerts its transforming potential by mimicking the growth promoting activity of PDGF and stimulating the cell in an autocrine manner. PDGF A-chain precursor polypeptide is assigned to the proximal long arm of chromosome 7, band q11.23. The human homolog(PDGF B-chain/c-sis) of the transforming gene of simian sarcoma virus is assigned to chromosome 22. The standard product used in this kit is recombinant human PDGF-AB with the molecular mass of 27KDa.

Synonyms: Platelet-derived growth factor subunit B,PDGF subunit B,PDGF-2,Platelet-derived growth factor B chain,Platelet-derived growth factor beta polypeptide,Proto-oncogene c-Sis,Becaplermin,PDGFB,PDGF2, SIS,

Full Gene Name: Platelet-derived growth factor subunit B

Cellular Localisation: Secreted. Released by platelets upon wounding.

Gene ID: 5155, 5154

UniProt: P01127