PDGF-AB Heterodimer ELISA Kit

Catalog No. ABIN1672747

Product Details

Target: PDGF-AB Heterodimer

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

Reactivity: Mouse

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 Mouse 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: <3pg/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:

Tissue Specificity: Localized to vascular smooth muscle cells. Also weakly expressed by cortical interstitial cells but absent in tubules. Up-regulated in areas of renal fibrosis. In mice with unilateral ureteral obstruction, an increased expression in interstitial cells and in some tubules observed after day 4. .

Plate: Pre-coated

Protocol: mouse PDGF-AB ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat 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 mouse 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 mouse 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 mouse cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each mouse PDGF-AB standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 262, Standard deviation: 12.3, CV(%): 4.7
• Sample 2: n=16, Mean(pg/ml): 524, Standard deviation: 27.8, CV(%): 5.3
• Sample 3: n=16, Mean(pg/ml): 924, Standard deviation: 57.3, CV(%): 6.2,
• Sample 1: n=24, Mean(pg/ml): 280, Standard deviation: 15.4, CV(%): 5.5
• Sample 2: n=24, Mean(pg/ml): 649, Standard deviation: 41.54, CV(%): 6.4
• Sample 3: n=24, Mean(pg/ml): 1107, Standard deviation: 80.8, CV(%): 7.3

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

Fernandes, Wang, Yuan, Liu, Dziak, Yang: "Combination of Controlled Release Platelet-Rich Plasma Alginate Beads and Bone Morphogenetic Protein-2 Genetically Modified Mesenchymal Stem Cells for Bone Regeneration." in: Journal of periodontology, Vol. 87, Issue 4, pp. 470-80, (2017) (PubMed).

Chancharoenthana, Leelahavanichkul, Taratummarat, Wongphom, Tiranathanagul, Eiam-Ong: "Cilostazol attenuates intimal hyperplasia in a mouse model of chronic kidney disease." in: PLoS ONE, Vol. 12, Issue 12, pp. e0187872, (2017) (PubMed).

Ge, Yu, Liu, Cong, Liu, Wang, Zhou, Lin: "Characterization of bone marrow-derived mesenchymal stem cells from dimethyloxallyl glycine-preconditioned mice: Evaluation of the feasibility of dimethyloxallyl glycine as a mobilization agent." in: Molecular medicine reports, Vol. 13, Issue 4, pp. 3498-506, (2016) (PubMed).

Target Details

Target: PDGF-AB Heterodimer

Alternative Name: 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. .

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.

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,Pdgfb,Sis,

Full Gene Name: Platelet-derived growth factor subunit B

Cellular Localisation: Secreted. Released by platelets upon wounding..

Gene ID: 18591, 18590

UniProt: P31240