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Human ADAM28 ELISA Kit

SKU:
HUFI01341
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q9UKQ2
Sensitivity:
9.375pg/ml
Range:
15.625-1000pg/ml
ELISA Type:
Sandwich
Synonyms:
ADAM28, Disintegrin and metalloproteinase domain-containing protein 28, ADAM 28, Epididymial metalloproteinase-like, disintegrin-like, and cysteine-rich protein II, eMDC II, Metalloproteinase-like, disintegrin-like, and cysteine-rich protein L, MDC-L
Reactivity:
Human
Research Area:
Cell Biology
€599
Frequently bought together:

Description

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Human ADAM28 ELISA Kit

The Human ADAM28 ELISA Kit is specifically designed for the precise measurement of ADAM28 levels in human serum, plasma, and cell culture supernatants. This kit offers exceptional sensitivity and specificity, guaranteeing accurate and consistent results, perfect for various research experiments.ADAM28, a member of the ADAM family of proteins, is a crucial enzyme involved in cell signaling, migration, and inflammation processes. Its dysregulation has been linked to various diseases, including cancer and inflammatory disorders.

Therefore, measuring ADAM28 levels can provide valuable insights into the pathogenesis of these conditions and aid in the development of potential therapeutic interventions.Don't miss out on the opportunity to study the role of ADAM28 in disease pathology with the Human ADAM28 ELISA Kit. Order yours today from www.assaygenie.com/human-adam28-elisa-kit/.

Product Name:Human ADAM28 ELISA Kit
Product Code:HUFI01341
Size:96 Assays
Alias:ADAM28, Disintegrin and metalloproteinase domain-containing protein 28, ADAM 28, Epididymial metalloproteinase-like, disintegrin-like, and cysteine-rich protein II, eMDC II, Metalloproteinase-like, disintegrin-like, and cysteine-rich protein L, MDC-L, ADAM23, MDCL, MDC-Lm, MDC-Ls, eMDCII
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human ADAM28 concentrations in serum plasma and other biological fluids.
Sensitivity:9.375pg/ml
Range:15.625-1000pg/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human ADAM28 and the recovery rates were calculated by comparing the measured value to the expected amount of Human ADAM28 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)92-10598
EDTA plasma(n=5)85-9991
UFH plasma(n=5)86-10391
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human ADAM28 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)86-105%86-96%85-102%
EDTA plasma(n=5)83-96%82-100%85-101%
UFH plasma(n=5)81-92%81-97%80-84%
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
UniprotQ9UKQ2
UniProt Protein Function:ADAM28: May play a role in the adhesive and proteolytic events that occur during lymphocyte emigration or may function in ectodomain shedding of lymphocyte surface target proteins, such as FASL and CD40L. May be involved in sperm maturation. 2 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:Protein type:EC 3.4.24.-; Membrane protein, integral; Motility/polarity/chemotaxis; Protease

Chromosomal Location of Human Ortholog: 8p21.2

Molecular Function:metallopeptidase activity

Biological Process: spermatogenesis

NCBI Summary:This gene encodes a member of the ADAM (a disintegrin and metalloprotease domain) family. Members of this family are membrane-anchored proteins structurally related to snake venom disintegrins, and have been implicated in a variety of biological processes involving cell-cell and cell-matrix interactions, including fertilization, muscle development, and neurogenesis. The protein encoded by this gene is a lymphocyte-expressed ADAM protein. This gene is present in a gene cluster with other members of the ADAM family on chromosome 8. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Jan 2015]
UniProt Code:Q9UKQ2
NCBI GenInfo Identifier:748983380
NCBI Gene ID:10863
NCBI Accession:NP_001291280.1
UniProt Related Accession:Q9UKQ2
Molecular Weight:
NCBI Full Name:disintegrin and metalloproteinase domain-containing protein 28 isoform 4 preproprotein
NCBI Synonym Full Names:ADAM metallopeptidase domain 28
NCBI Official Symbol:ADAM28  
NCBI Official Synonym Symbols:MDCL; MDC-L; eMDCII; ADAM 28; eMDC II  
NCBI Protein Information:disintegrin and metalloproteinase domain-containing protein 28
UniProt Protein Name:Disintegrin and metalloproteinase domain-containing protein 28
UniProt Synonym Protein Names:Epididymal metalloproteinase-like, disintegrin-like, and cysteine-rich protein II; eMDC II; Metalloproteinase-like, disintegrin-like, and cysteine-rich protein L; MDC-L
UniProt Gene Name:ADAM28  

*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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