PDGF-AB Heterodimer ELISA Kit

Catalog No. ABIN1112778

Product Details

Target: PDGF-AB Heterodimer

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 31.2-2000 pg/mL

Minimum Detection Limit: 31.2 pg/mL

Application: ELISA

Analytical Method: Quantitative

Sensitivity: < 3 pg/mL

Components: 1. One 96-well plate pre-coated with anti-Rat PDGF-AB antibody 2. Lyophilized Rat PDGF-AB standards: 2 tubes (10ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-Rat PDGF-AB antibody (Concentrated): 130 µl.

Material not included: 1. 37 °C incubator 2. Microplate reader (wavelength: 450nm) 3. Precise pipette and disposable pipette tips 4. Automated plate washer 5. ELISA shaker 6. 1.5ml of Eppendorf tubes 7. Plate cover 8. Absorbent filter papers 9. Plastic or glass container with volume of above 1L

Application Details

Comment:

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-PDGF-AB polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-PDGF-AB polyclonal antibody was used as detection antibodies. The standards test samples and biotin conjugated detection antibody were added - the wells subsequently and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used - visualize HRP enzymatic reaction. TMB was catalyzed by HRP - produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional - the PDGF-AB amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of PDGF-AB can be calculated.

Plate: Pre-coated

Reagent Preparation:

1. Before the experiment, centrifuge each kit component for several minutes to bring down all reagents to the bottom of tubes. 2. It is recommend to measure each standard and sample in duplicate. 3. Do NOT let the plate completely dry at any time! Since the dry condition can inactivate the biological material on the plate. 4. Do not reuse pipette tips and tubes to avoid cross contamination. 5. Do not use the expired components and the components from different batches. 6. To avoid the marginal effect of plate incubation for temperature differences (the marginal wells always get stronger reaction), it is recommend to equilibrate the ABC working solution and TMB substrate for at least 30 min at room temperature (37°C ) before adding to wells.The TMB substrate (Kit Component 8) is colorless and transparent before use, if not, please contact us for replacement.

Sample Preparation:

Preparation of sample and reagents 1. Sample Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20 °C for long term. Avoid multiple freeze-thaw cycles.
Tissue lysate, body fluids and cell culture supernatants: Centrifuge to remove precipitate, analyze immediately or aliquot and store at -20 °C .
Serum: Coagulate the serum at room temperature (about 4 hours). Centrifuge at approximately 1500 × g for 15 min. Analyze the serum immediately or aliquot and store at -20 °C .
Plasma: Collect plasma with heparin or EDTA as the anticoagulant. Centrifuge for 15 min at 1500 x g within 30 min of collection. For eliminating platelet, suggesting that further centrifugation for 10 min at 2-8°C at 10000 x g. Analyze immediately or aliquot and store frozen at -20 °C. Analyze immediately or aliquot and store frozen at -20 °C. Citrate can not be used as anticoagulant here. Note: 1. Coagulate blood samples completely, then, centrifuge, and avoid hemolysis and particle. 2. NaN3 can not be used as test sample preservative, since it is the inhibitor for HRP.

Restrictions: For Research Use only

Handling

Preservative: Sodium azide, Thimerosal (Merthiolate)

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: Platelet-derived growth factor (PDGF) was one of the first growth factors characterized. and also a potent mitogen for cells of mesenchymal origin, including smooth muscle cells and glial cells. It regulates cell growth and division, In particular, it plays a significant role in blood vessel formation (angiogenesis). It is produced by a plethora of cells, and synthesized, stored, released by platelets upon activation. In chemical terms, PSGF is dimeric glycoprotein composed of two A (-AA) or two B (-BB) chains or a combination of the two (-AB). It is found that initial PDGF-AB interaction with the alpha PDGFR induces conformational changes in the ligand or receptor that facilitates efficient recruitment of beta PDGFR by this PDGF isoform.