WB
Western blot analysis of SIRT1 on different lysates with Rabbit anti-SIRT1 antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate (15 µg/Lane) Lane 2: HEK-293 cell lysate (15 µg/Lane) Lane 3: A549 cell lysate (15 µg/Lane) Lane 4: F9 cell lysate (15 µg/Lane) Lane 5: Mouse testis tissue lysate (20 µg/Lane) Lane 6: Rat testis tissue lysate (20 µg/Lane) Exposure time: 2 minutes 37 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-SIRT1 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of HEK-293 cells labeling SIRT1 with Rabbit anti-SIRT1 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SIRT1 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either SIRT1 or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.| Product Name | SIRT1 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human SIRT1 aa 698-747 / 747. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ChIPICC/IFIHCWB |
| WB:1:2000 IHC:1:50-1:1000 ICC:1:50-1:200 ChIP:Use 0.5~2μg for 25μg of chromatin. |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | SIRT1 |
|---|---|
| Gene Synonyms | SIR2 SIR2L1 SIR2alpha |
| Gene Full Name | sirtuin 1 |
| Gene 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 I of the sirtuin family. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Dec 2008] |
| Alternative Names | 75SirT1 antibody hSIR2 antibody hSIRT1 antibody HST2 S. cerevisiae homolog of antibody NAD dependent deacetylase sirtuin 1 antibody NAD dependent protein deacetylase sirtuin 1 antibody OTTHUMP00000198111 antibody OTTHUMP00000198112 antibody Regulatory protein SIR2 homolog 1 antibody SIR1_HUMAN antibody 75SirT1 antibody hSIR2 antibody hSIRT1 antibody HST2 S. cerevisiae homolog of antibody NAD dependent deacetylase sirtuin 1 antibody NAD dependent protein deacetylase sirtuin 1 antibody OTTHUMP00000198111 antibody OTTHUMP00000198112 antibody Regulatory protein SIR2 homolog 1 antibody SIR1_HUMAN antibody SIR2 antibody SIR2 like 1 antibody SIR2 like protein 1 antibody SIR2 S.cerevisiae homolog-like 1 antibody SIR2-like protein 1 antibody SIR2ALPHA antibody SIR2L1 antibody Sirt1 antibody SirtT1 75 kDa fragment antibody Sirtuin (silent mating type information regulation 2 homolog) 1 (S. cerevisiae) antibody Sirtuin 1 antibody Sirtuin type 1 antibody |
| Molecular Weight(MW) | 82kDa(Observed band size: 110kDa) |
| Cellular Localization | Nucleus, Cytoplasm, Mitochondrion. |
WB
Western blot analysis of SIRT1 on different lysates with Rabbit anti-SIRT1 antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate (15 µg/Lane) Lane 2: HEK-293 cell lysate (15 µg/Lane) Lane 3: A549 cell lysate (15 µg/Lane) Lane 4: F9 cell lysate (15 µg/Lane) Lane 5: Mouse testis tissue lysate (20 µg/Lane) Lane 6: Rat testis tissue lysate (20 µg/Lane) Exposure time: 2 minutes 37 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-SIRT1 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of HEK-293 cells labeling SIRT1 with Rabbit anti-SIRT1 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SIRT1 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either SIRT1 or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.| Application Notes | WB:1:2000 IHC:1:50-1:1000 ICC:1:50-1:200 ChIP:Use 0.5~2μg for 25μg of chromatin. |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
