Human IGFBP3 ELISA Kit (HUFI00152)- High Sensitivity - Assay Genie

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Human IGFBP3 ELISA Kit (HUFI00152)

Rating Required Select Rating 1 star (worst) 2 stars 3 stars (average) 4 stars 5 stars (best) Name Review Subject Required Comments Required SKU: HUFI00152 UPC: Product Type: ELISA Kit Size: 96 Assays Uniprot: P17936 Sensitivity: 0.234ng/ml Range: 0.391-25ng/ml ELISA Type: Sandwich Synonyms: IGFBP-3, IGFBP3, BP-53, IBP3 Reactivity: Human

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Description

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Human IGFBP3 ELISA Kit

The Human IGFBP3 (Insulin-like Growth Factor Binding Protein 3) ELISA Kit is specifically designed for the accurate quantification of IGFBP3 levels in human serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit ensures reliable and reproducible results for a variety of research applications.IGFBP3 is a critical binding protein that regulates the bioavailability of insulin-like growth factors, impacting cell growth, differentiation, and survival. Dysregulation of IGFBP3 has been linked to various diseases, including cancer, diabetes, and cardiovascular disorders, highlighting its importance as a biomarker for disease progression and therapeutic development.

This ELISA kit provides a convenient and efficient method for studying IGFBP3 levels in biological samples, offering researchers valuable insights into disease mechanisms and potential treatment strategies. Trust in the Human IGFBP3 ELISA Kit for accurate and precise measurements, advancing your research endeavors in the field of molecular biology and medicine.

Overview

Product Name:	Human IGFBP3 ELISA Kit
Product Code:	HUFI00152
Size:	96 Assays
Alias:	IGFBP-3, IGFBP3, BP-53, IBP3, Insulin-like growth factor-binding protein 3
Detection method:	Sandwich ELISA, Double Antibody
Application:	This immunoassay kit allows for the in vitro quantitative determination of Human IGFBP-3 concentrations in serum plasma and other biological fluids.
Sensitivity:	0.234ng/ml
Range:	0.391-25ng/ml
Storage:	4°C for 6 months
Note:	For Research Use Only

Additional Information

Recovery:	Matrices listed below were spiked with certain level of Human IGFBP-3 and the recovery rates were calculated by comparing the measured value to the expected amount of Human IGFBP-3 in samples.
Matrix Recovery range(%) Average(%) serum(n=5) 85-103 92 EDTA plasma(n=5) 87-105 98 UFH plasma(n=5) 85-97 93
Linearity:	The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human IGFBP-3 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample 1:2 1:4 1:8 serum(n=5) 87-99% 85-96% 86-101% EDTA plasma(n=5) 87-98% 86-98% 85-93% UFH plasma(n=5) 82-88% 85-100% 81-96%
CV(%):	Intra-Assay: CV<8% Inter-Assay: CV<10%

Kit Components

Component	Quantity	Storage
ELISA Microplate (Dismountable)	8×12 strips	4°C for 6 months
Lyophilized Standard	2	4°C/-20°C
Sample/Standard Dilution Buffer	20ml	4°C
Biotin-labeled Antibody(Concentrated)	120ul	4°C (Protect from light)
Antibody Dilution Buffer	10ml	4°C
HRP-Streptavidin Conjugate(SABC)	120ul	4°C (Protect from light)
SABC Dilution Buffer	10ml	4°C
TMB Substrate	10ml	4°C (Protect from light)
Stop Solution	10ml	4°C
Wash Buffer(25X)	30ml	4°C
Plate Sealer	5	-

Other materials and equipment required:

• Microplate reader with 450 nm wavelength filter
• Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
• Incubator
• Deionized or distilled water
• Absorbent paper
• Buffer resevoir

Protein Information

Uniprot

P17936

UniProt Protein Function:	IGFBP3: IGF-binding proteins prolong the half-life of the IGFs and have been shown to either inhibit or stimulate the growth promoting effects of the IGFs on cell culture. They alter the interaction of IGFs with their cell surface receptors. Also exhibits IGF-independent antiproliferative and apoptotic effects mediated by its receptor TMEM219/IGFBP-3R.
UniProt Protein Details:	Protein type:Secreted; Cell development/differentiation; Secreted, signal peptide Chromosomal Location of Human Ortholog: 7p12.3 Cellular Component: extracellular region; extracellular space; insulin-like growth factor binding protein complex; nucleus Molecular Function:fibronectin binding; insulin-like growth factor binding; insulin-like growth factor I binding; insulin-like growth factor II binding; metal ion binding; protein binding; protein tyrosine phosphatase activator activity Biological Process: apoptosis; cellular protein metabolic process; negative regulation of cell proliferation; negative regulation of protein amino acid phosphorylation; negative regulation of signal transduction; negative regulation of smooth muscle cell migration; negative regulation of smooth muscle cell proliferation; osteoblast differentiation; positive regulation of apoptosis; positive regulation of catalytic activity; positive regulation of insulin-like growth factor receptor signaling pathway; positive regulation of MAPKKK cascade; positive regulation of myoblast differentiation; protein amino acid phosphorylation; regulation of cell growth; regulation of insulin-like growth factor receptor signaling pathway
NCBI Summary:	This gene is a member of the insulin-like growth factor binding protein (IGFBP) family and encodes a protein with an IGFBP domain and a thyroglobulin type-I domain. The protein forms a ternary complex with insulin-like growth factor acid-labile subunit (IGFALS) and either insulin-like growth factor (IGF) I or II. In this form, it circulates in the plasma, prolonging the half-life of IGFs and altering their interaction with cell surface receptors. Alternate transcriptional splice variants, encoding different isoforms, have been characterized. [provided by RefSeq, Jul 2008]
UniProt Code:	P17936
NCBI GenInfo Identifier:	146327827
NCBI Gene ID:	3486
NCBI Accession:	P17936.2
UniProt Secondary Accession:	P17936,Q2V509, Q6P1M6, Q9UCL4, A4D2F5, D3DVM0,
UniProt Related Accession:	P17936
Molecular Weight:	32,223 Da
NCBI Full Name:	Insulin-like growth factor-binding protein 3
NCBI Synonym Full Names:	insulin like growth factor binding protein 3
NCBI Official Symbol:	IGFBP3
NCBI Official Synonym Symbols:	IBP3; BP-53
NCBI Protein Information:	insulin-like growth factor-binding protein 3
UniProt Protein Name:	Insulin-like growth factor-binding protein 3
Protein Family:	Insulin-like growth factor-binding protein
UniProt Gene Name:	IGFBP3
UniProt Entry Name:	IBP3_HUMAN

Protocol

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

Step	Protocol
1.	Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.	Aliquot 0.1ml standard solutions into the standard wells.
3.	Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.	Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.	Seal the plate with a cover and incubate at 37 °C for 90 min.
6.	Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.	Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.	Seal the plate with a cover and incubate at 37°C for 60 min.
9.	Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.	Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11.	Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.	Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.	Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14.	Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

Sample Preparation

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type	Protocol
Serum	If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.
Plasma	Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal Fluid	Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatant	Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysates	Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenates	The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysates	Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast Milk	Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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