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Human ITGB3 / CD61 / Integrin beta-3 ELISA Kit

SKU:
HUFI01455
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P05106
Sensitivity:
46.875pg/ml
Range:
78.125-5000pg/ml
ELISA Type:
Sandwich
Synonyms:
ITGB3, Integrin beta-3, Platelet membrane glycoprotein IIIa, GPIIIa, GP3A, CD61, INGRB3, ITG B3
Reactivity:
Human
Research Area:
Cell Biology
$719
Frequently bought together:

Description

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Human ITGB3/CD61/Integrin beta-3 ELISA Kit

The Human ITGB3 (CD61 Integrin Beta-3) ELISA Kit is specifically designed for the accurate detection of ITGB3 levels in human serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring precise and consistent results for various research applications.Integrin Beta-3, also known as CD61, is a key protein involved in cell adhesion and signaling processes. It plays a crucial role in various physiological and pathological processes, including cell migration, wound healing, and cancer metastasis.

As a result, ITGB3 is a valuable biomarker for studying these processes and developing potential therapeutic interventions.With this ELISA kit, researchers can confidently measure ITGB3 levels in biological samples, providing valuable insights into cellular pathways and disease mechanisms. Its ease of use, accuracy, and reliability make it an essential tool for investigating integrin beta-3 biology and its implications in health and disease.

Product Name:Human ITGB3 / CD61 / Integrin beta-3 ELISA Kit
Product Code:HUFI01455
Size:96 Assays
Alias:ITGB3, Integrin beta-3, Platelet membrane glycoprotein IIIa, GPIIIa, GP3A, CD61, INGRB3, ITG B3
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human ITGB3 concentrations in serum plasma and other biological fluids.
Sensitivity:46.875pg/ml
Range:78.125-5000pg/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human ITGB3 and the recovery rates were calculated by comparing the measured value to the expected amount of Human ITGB3 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)88-10594
EDTA plasma(n=5)86-9890
UFH plasma(n=5)87-10594
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human ITGB3 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)85-96%86-104%85-98%
EDTA plasma(n=5)84-101%89-97%84-101%
UFH plasma(n=5)80-96%81-95%82-96%
CV(%):Intra-Assay: CV<8%
Inter-Assay: CV<10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

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
UniprotP05106
UniProt Protein Function:ITGB3: integrin alpha-V/beta-3 is a receptor for cytotactin, fibronectin, laminin, matrix metalloproteinase-2, osteopontin, osteomodulin, prothrombin, thrombospondin, vitronectin and von Willebrand factor. Integrin alpha-IIB/beta-3 is a receptor for fibronectin, fibrinogen, plasminogen, prothrombin, thrombospondin and vitronectin. Integrins alpha-IIB/beta-3 and alpha-V/beta-3 recognize the sequence R-G-D in a wide array of ligands. Integrin alpha-IIB/beta-3 recognizes the sequence H-H-L-G-G-G-A-K-Q-A-G-D-V in fibrinogen gamma chain. Following activation integrin alpha- IIB/beta-3 brings about platelet/platelet interaction through binding of soluble fibrinogen. This step leads to rapid platelet aggregation which physically plugs ruptured endothelial surface.
UniProt Protein Details:Protein type:Membrane protein, integral; Motility/polarity/chemotaxis; Cell adhesion

Chromosomal Location of Human Ortholog: 17q21.32

Cellular Component: cell surface; filopodium membrane; focal adhesion; integral to plasma membrane; integrin complex; melanosome; microvillus membrane; nucleus; plasma membrane; platelet alpha granule membrane; receptor complex

Molecular Function:C-X3-C chemokine binding; cell adhesion molecule binding; coreceptor activity; enzyme binding; extracellular matrix binding; fibroblast growth factor binding; fibronectin binding; identical protein binding; insulin-like growth factor I binding; platelet-derived growth factor receptor binding; protease binding; protein binding; protein disulfide isomerase activity; vascular endothelial growth factor receptor 2 binding

Biological Process: activation of protein kinase activity; angiogenesis involved in wound healing; blood coagulation; cell adhesion; cell growth; cell migration; cell-matrix adhesion; cell-substrate adhesion; entry of virus into host cell; extracellular matrix organization and biogenesis; heterotypic cell-cell adhesion; integrin-mediated signaling pathway; leukocyte migration; mesodermal cell differentiation; negative chemotaxis; negative regulation of lipid transport; negative regulation of lipoprotein metabolic process; negative regulation of low-density lipoprotein receptor biosynthetic process; platelet activation; platelet degranulation; positive regulation of endothelial cell proliferation; positive regulation of peptidyl-tyrosine phosphorylation; positive regulation of protein amino acid phosphorylation; positive regulation of vascular endothelial growth factor receptor signaling pathway; regulation of bone resorption; smooth muscle cell migration; tube development; vascular endothelial growth factor receptor signaling pathway; wound healing

Disease: Bleeding Disorder, Platelet-type, 16; Glanzmann Thrombasthenia; Myocardial Infarction, Susceptibility To

NCBI Summary:The ITGB3 protein product is the integrin beta chain beta 3. Integrins are integral cell-surface proteins composed of an alpha chain and a beta chain. A given chain may combine with multiple partners resulting in different integrins. Integrin beta 3 is found along with the alpha IIb chain in platelets. Integrins are known to participate in cell adhesion as well as cell-surface mediated signalling. [provided by RefSeq, Jul 2008]
UniProt Code:P05106
NCBI GenInfo Identifier:125987835
NCBI Gene ID:3690
NCBI Accession:P05106.2
UniProt Secondary Accession:P05106,O15495, Q12806, Q13413, Q14648, Q16499, A0PJW2 D3DXJ8,
UniProt Related Accession:P05106
Molecular Weight:86,694 Da
NCBI Full Name:Integrin beta-3
NCBI Synonym Full Names:integrin subunit beta 3
NCBI Official Symbol:ITGB3
NCBI Official Synonym Symbols:GT; CD61; GP3A; BDPLT2; GPIIIa; BDPLT16
NCBI Protein Information:integrin beta-3
UniProt Protein Name:Integrin beta-3
UniProt Synonym Protein Names:Platelet membrane glycoprotein IIIa; GPIIIa; CD_antigen: CD61
Protein Family:Integrin
UniProt Gene Name:ITGB3
UniProt Entry Name:ITB3_HUMAN

*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.

StepProtocol
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.

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 TypeProtocol
SerumIf 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.

PlasmaCollect 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 FluidCollect 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 supernatantCollect 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 lysatesSolubilize 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 homogenatesThe 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 lysatesRinse 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 MilkCollect 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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