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Human SIRT7 ELISA Kit

SKU:
HUFI01031
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q9NRC8
Sensitivity:
0.188ng/ml
Range:
0.313-20ng/ml
ELISA Type:
Sandwich
Synonyms:
SIRT7, NAD-dependent deacetylase sirtuin-7, SIR2L7, Regulatory protein SIR2 homolog 7, SIR2-like protein 7
Reactivity:
Human
Research Area:
Epigenetics and Nuclear Signaling
€599
Frequently bought together:

Description

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Human SIRT7 ELISA Kit

The Human SIRT7 ELISA Kit is a cutting-edge tool for the precise measurement of SIRT7 levels in human samples, including serum, plasma, and cell culture supernatants. With its exceptional sensitivity and specificity, this kit delivers accurate and consistent results, making it an invaluable resource for various research endeavors.SIRT7 is a key enzyme belonging to the sirtuin family, known for its involvement in regulating various cellular processes such as gene expression, metabolism, and aging. It plays a critical role in maintaining cellular homeostasis and has been implicated in various diseases, including cancer, metabolic disorders, and neurodegenerative conditions, making it a promising target for therapeutic development.

By utilizing the Human SIRT7 ELISA Kit, researchers can gain valuable insights into the function and expression of SIRT7 in different biological contexts, paving the way for the discovery of novel therapeutic strategies and diagnostic markers. Choose this advanced assay kit to uncover the mysteries of SIRT7 and advance your scientific exploration.

Product Name:Human SIRT7 ELISA Kit
Product Code:HUFI01031
Size:96 Assays
Alias:SIRT7, NAD-dependent deacetylase sirtuin-7, SIR2L7, Regulatory protein SIR2 homolog 7, SIR2-like protein 7
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human SIRT7 concentrations in serum plasma and other biological fluids.
Sensitivity:0.188ng/ml
Range:0.313-20ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human SIRT7 and the recovery rates were calculated by comparing the measured value to the expected amount of Human SIRT7 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)86-10092
EDTA plasma(n=5)86-10593
UFH plasma(n=5)96-10398
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human SIRT7 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)85-102%97-105%85-101%
EDTA plasma(n=5)84-99%90-101%84-99%
UFH plasma(n=5)80-92%80-99%88-95%
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
UniprotQ9NRC8
UniProt Protein Function:SIRT7: NAD-dependent protein deacetylase that specifically mediates deacetylation of histone H3 at 'Lys-18' (H3K18Ac). In contrast to other histone deacetylases, displays selectivity for a single histone mark, H3K18Ac, directly linked to control of gene expression. H3K18Ac is mainly present around the transcription start site of genes and has been linked to activation of nuclear hormone receptors. SIRT7 thereby acts as a transcription repressor. Moreover, H3K18 hypoacetylation has been reported as a marker of malignancy in various cancers and seems to maintain the transformed phenotype of cancer cells. These data suggest that SIRT7 may play a key role in oncogenic transformation by suppresses expression of tumor suppressor genes by locus-specific deacetylation of H3K18Ac at promoter regions. Also required to restore the transcription of ribosomal RNA (rRNA) at the exit from mitosis: promotes the association of RNA polymerase I with the rDNA promoter region and coding region. Stimulates transcription activity of the RNA polymerase I complex. May also deacetylate p53/TP53 and promotes cell survival, however such data need additional confirmation. Belongs to the sirtuin family. Class IV subfamily. 3 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:Protein type:Nucleolus; EC 3.5.1.-; Deacetylase

Chromosomal Location of Human Ortholog: 17q25

Cellular Component: nucleolus; nucleolus organizer region

Molecular Function:chromatin binding; protein binding

Biological Process: activation of transcription on exit from mitosis; negative regulation of transcription from RNA polymerase II promoter; rRNA transcription

NCBI Summary:This gene encodes a member of the sirtuin family of proteins, homologs to the yeast Sir2 protein. Members of the sirtuin family are characterized by a sirtuin core domain and grouped into four classes. The functions of human sirtuins have not yet been determined; however, yeast sirtuin proteins are known to regulate epigenetic gene silencing and suppress recombination of rDNA. Studies suggest that the human sirtuins may function as intracellular regulatory proteins with mono-ADP-ribosyltransferase activity. The protein encoded by this gene is included in class IV of the sirtuin family. [provided by RefSeq, Jul 2008]
UniProt Code:Q9NRC8
NCBI GenInfo Identifier:38258650
NCBI Gene ID:51547
NCBI Accession:Q9NRC8.1
UniProt Secondary Accession:Q9NRC8,Q3MIK4, Q9NSZ6, Q9NUS6, A8K2K0, B3KSU8,
UniProt Related Accession:Q9NRC8
Molecular Weight:35,851 Da
NCBI Full Name:NAD-dependent protein deacetylase sirtuin-7
NCBI Synonym Full Names:sirtuin 7
NCBI Official Symbol:SIRT7
NCBI Official Synonym Symbols:SIR2L7
NCBI Protein Information:NAD-dependent protein deacetylase sirtuin-7
UniProt Protein Name:NAD-dependent protein deacetylase sirtuin-7
UniProt Synonym Protein Names:Regulatory protein SIR2 homolog 7; SIR2-like protein 7
Protein Family:NAD-dependent protein deacetylase
UniProt Gene Name:SIRT7
UniProt Entry Name:SIR7_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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