PDGF-AB Heterodimer ELISA Kit

Catalog No. ABIN411346

Rat ELISA Kit Colorimetric assay for quantification of Rat .

Quick Overview for PDGF-AB Heterodimer ELISA Kit (ABIN411346)

Target: PDGF-AB Heterodimer

Binding Specificity: AA 74-182, AA 86-204

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 31.2-2000 pg/mL

Application: ELISA

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

Product Details

Minimum Detection Limit: 31.2 pg/mL

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

Brand: PicoKine™

Analytical Method: Quantitative

Specificity: Expression system for standard: E.coli
Immunogen sequence: (R86-K204)+(S74-T182)

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: (R86-K204)+(S74-T182)

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.

Plate: Pre-coated

Protocol: rat 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, (R86-K204)+(S74-T182)) 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 rat 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 rat 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 rat cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each rat PDGF-AB standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 246, Standard deviation: 14.27, CV(%): 5.8
• Sample 2: n=16, Mean(pg/ml): 632, Standard deviation: 34.76, CV(%): 5.5
• Sample 3: n=16, Mean(pg/ml): 1068, Standard deviation: 50.2, CV(%): 4.7,
• Sample 1: n=24, Mean(pg/ml): 275, Standard deviation: 20.35, CV(%): 7.4
• Sample 2: n=24, Mean(pg/ml): 723, Standard deviation: 45.55, CV(%): 6.3
• Sample 3: n=24, Mean(pg/ml): 1249, Standard deviation: 73.7, CV(%): 5.9

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

Target Details

Target: PDGF-AB Heterodimer

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

Background:

Protein Function: Pdgfa: Growth factor that plays an essential role in theregulation of embryonic development, cell proliferation, cellmigration, survival and chemotaxis. Potent mitogen for cells ofmesenchymal origin. Required for normal lung alveolar septumformation during embryogenesis, normal development of thegastrointestinal tract, normal development of Leydig cells andspermatogenesis. Required for normal oligodendrocyte developmentand normal myelination in the spinal cord and cerebellum. Plays animportant role in wound healing. Signaling is modulated by theformation of heterodimers with PDGFB (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 rat PDGF-AB(A chain Ser87-Arg196, B chain Ser74-Thr182) with the molecular mass of 24.8KDa.

Synonyms: Platelet derived growth factor, B polypeptide ,Platelet-derived growth factor subunit B ,Pdgfb ,rCG_59494 ,

Full Gene Name: platelet-derived growth factor beta polypeptide,platelet-derived growth factor alpha polypeptide

Cellular Localisation: Pdgfa: Secreted. Note: Released by platelets uponwounding.

Gene ID: 24628, 25266

UniProt: G3V882