IGFBPI ELISA Kit

Catalog No. ABIN1112632

Human IGFBPI ELISA Kit Colorimetric assay for quantification of Human IGFBPI in Cell Culture Supernatant, Plasma, Serum and Tissue Lysate

Quick Overview for IGFBPI ELISA Kit (ABIN1112632)

Target: IGFBPI (Insulin-Like Growth Factor Binding Protein 1 (IGFBPI))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 31.2-2000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Plasma, Serum, Tissue Lysate

Product Details IGFBPI ELISA Kit

Minimum Detection Limit: 31.2 pg/mL

Purpose: For quantitative detection of IGFBP-1 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Analytical Method: Quantitative

Sensitivity: < 1 pg/mL

Components: 1. One 96-well plate pre-coated with anti-Human IGFBP-1 antibody 2. Lyophilized Human IGFBP-1 standards: 2 tubes (10ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-Human IGFBP-1 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-IGFBP-1 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-IGFBP-1 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 IGFBP-1 amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of IGFBP-1 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.
Body fluids, tissue lysates 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 1000 × g for 15 min. Analyze the serum immediately or aliquot and store at -20 °C .
Plasma: Collect plasma on ice cube with heparin or EDTA as the anticoagulant. Centrifuge for 20 min at 2000 x g within 30 min of collection. 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 for IGFBPI

Target: IGFBPI (Insulin-Like Growth Factor Binding Protein 1 (IGFBPI))

Alternative Name: IGFBP-1

Background: Insulin-like growth factor-binding protein 1 (IGFBP-1) also known as placental protein 12 (PP12) is a member of the Insulin-like growth factor-binding protein (IGFBP) family. It is synthesized in liver, secretory endometrium, and decidua. The IGFBP1 gene has 4 exons, spans 5.9 kb, and mapped to 7p13-p12. This protein has an IGFBP domain and a type-I thyroglobulin domain. It binds both insulin-like growth factors (IGFs) I and II and circulates in the plasma. Binding of this protein prolongs the half-life of the IGFs and alters their interaction with cell surface receptors. Leu and George (2007) concluded that IGFBP1 is a negative regulator of the p53/BAK-dependent pathway of apoptosis.

Pathways: Myometrial Relaxation and Contraction, ER-Nucleus Signaling, Growth Factor Binding