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Product name
SIRT1 Human ELISA Kit
Specificity
Species– human. Rat, mouse are not suitable with this kit, other species are untested.
Precision
| Sample | n | Mean | SD | CV% |
|---|---|---|---|---|
| Overall | 8 | 7% |
| Sample | n | Mean | SD | CV% |
|---|---|---|---|---|
| Overall | 3 | 7.5% |
Tests
1 x 96 well plate
Sample type
Tissue Extracts, Cell Lysate
Assay type
Sandwich
Sensitivity
8 ng/ml
Recovery
| Sample type | Average % | Range % |
|---|---|---|
| Serum | 106 | 98 - 112 |
Cross reactivity
Reacts with
Human
Product overview
ab123457 SIRT1 Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human SIRT1 in cell and tissue lysates. The assay employs an antibody specific for Human SIRT1 protein coated onto well plate strips.
Standards and samples are pipetted into the wells and analyte present in the sample is bound to the wells by the immobilized antibody. The wells are washed and an anti- SIRT1 primary detector antibody is added. After washing away unbound primary detector antibody, HRP-conjugated secondary detector antibody specific for the primary detector antibody is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of analyte bound. The developing blue color is measured at 600 nm.
Optionally the reaction can be stopped by adding hydrochloric acid which changes the color from blue to yellow and the intensity can be measured at 450 nm.
Notes
Typical working ranges:
Tissue extract (e.g. Heart): None detected
Whole cultured cell extract: 0.060 - 1 mg/mL
Human SIRT1 standard: 0.008 - 0.5 µg/mL
Tissue extract (e.g. Heart): None detected
Whole cultured cell extract: 0.060 - 1 mg/mL
Human SIRT1 standard: 0.008 - 0.5 µg/mL
Store all components at 4°C. This kit is stable for at least 6 months from receipt. After reconstitution the standard should be stored at -80°C. Unused microplate strips should be returned to the pouch containing the desiccant and resealed.
Tested applications
Sandwich ELISAmore details
Properties
Storage instructions
Please see Notes section
| Components | Identifier | 1 x 96 tests |
|---|---|---|
| 10X HRP Label | 8206020 | 1 x 1ml |
| 10X Blocking Buffer | 8209802 | 1 x 6ml |
| ab110304 - 10X SIRT1 Detector Antibody | 8209540 | 1 x 1ml |
| 20X Buffer | 8203013 | 1 x 20ml |
| Extraction Buffer | 8201093 | 1 x 15ml |
| ab54334 - Human SIRT1 Standard | 8209542 | 1 x 1µg |
| SIRT1 Microplate | 8209541 | 1 unit |
| TMB Development Solution | 8209803 | 1 x 12ml |
Function
NAD-dependent protein deacetylase, which regulates processes such as apoptosis and muscle differentiation by deacetylating key proteins. Deacetylates 'Lys-382' of p53/TP53 and impairs its ability to induce proapoptotic program and modulate cell senescence. Deacetylates TAF1B and thereby represses rDNA transcription by the RNA polymerase I. Deacetylates 'Lys-266' of SUV39H1, leading to its activation. Deacetylates 'Lys-26' of HIST1H1E. Involved in HES1- and HEY2-mediated transcriptional repression. Inhibits skeletal muscle differentiation by deacetylating PCAF and MYOD1. May serve as a sensor of the cytosolic ratio of NAD(+)/NADH, which is essential in skeletal muscle cell differentiation. Deacetylates 'Lys-16' of histone H4 (in vitro). Component of the eNoSC (energy-dependent nucleolar silencing) complex, a complex that mediates silencing of rDNA in response to intracellular energy status and acts by recruiting histone-modifying enzymes. The eNoSC complex is able to sense the energy status of cell: upon glucose starvation, elevation of NAD(+)/NADP(+) ratio activates SIRT1, leading to histone H3 deacetylation followed by dimethylation of H3 at 'Lys-9' (H3K9me2) by SUV39H1 and the formation of silent chromatin in the rDNA locus. Deacetylates H2A. In case of HIV-1 infection, interacts with and deacetylates the viral Tat protein. Deacetylates APEX1 at 'Lys-6' and 'Lys-7'. Stimulates cellular AP endonuclease activity by promoting the association of APEX1 to XRCC1.
Tissue specificity
Widely expressed.
Sequence similarities
Belongs to the sirtuin family.
Contains 1 deacetylase sirtuin-type domain.
Post-translational
modifications
S-nitrosylated by GAPDH, leading to inhibit the NAD-dependent protein deacetylase activity.
Cellular localization
Nucleus > PML body. Recruited to the nuclear bodies via its interaction with PML. Colocalized with APEX1 in the nucleus. May be found in nucleolus, nuclear euchromatin, heterochromatin and inner membrane.
Target information above from: UniProt accessionQ96EB6
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- hSIR2
- hSIRT1
- NAD-dependent deacetylase sirtuin-1
see all
Database links
- Entrez Gene: 23411 Human
- SwissProt: Q96EB6 Human
- Unigene: 369779 Human
Applications
Show applications key
Our Abpromise guarantee covers the use of ab123457 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
sELISA: Use at an assay dependent concentration.
SIRT1 Human ELISA Kit images:
Sandwich ELISA - SIRT1 Human ELISA Kit (ab123457)
Example standard curve. Serially diluted recombinant SIRT1 in the working range of the assay.
Sandwich ELISA - SIRT1 Human ELISA Kit (ab123457)
Example of serially titrated HeLa extract in the assay working range.
Sandwich ELISA - SIRT1 Human ELISA Kit (ab123457)
The expression level of SIRT1 varies by cell type. Four cell types are analyzed by SIRT1 expression level relative to the Human HepG2 cell line. This relative relationship is confirmed by Western blotting using antibody ab13533.
Immunocytochemistry/ Immunofluorescence - SIRT1 Human ELISA Kit (ab123457)
The SIRT1 detector antibody (ab110304) in this ELISA is available as standalone antibody. In this image Immunocytochemistry of the antibody (ab110304) from this ELISA used to stain Human HDFn cells.
Protocols
References for SIRT1 Human ELISA Kit (ab123457)
ab123457 has not yet been referenced specifically in any publications.
Publishing research using ab123457? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"