ER130811

SIRT1 Rabbit Polyclonal Antibody

cat.: ER130811

Product Type:	Rabbit polyclonal IgG, primary antibodies
Species reactivity:	Human, Mouse
Applications:	WB, IHC-P, FC, IF-Cell
Clonality:	Polyclonal

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Immunogen affinity purified.
Molecular weight:	Predicted band size: 82 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human SIRT1 aa 698-747 / 747.
Positive control:	Hela cell lysate, Jurkat cell lysate, F9 cell lysate, HeLa, F9, human colon carcinoma tissue, human lung carcinoma tissue, mouse liver tissue, mouse testis tissue.
Subcellular location:	Nucleus, cytoplasm, Mitochondrion.
Recommended Dilutions: WB IHC-P FC IF-Cell	1:1,000-1:2,000 1:200 1:1,000 1:100
Uniprot #:	SwissProt: Q96EB6 Human
Alternative names:	75SirT1 hSIR2 hSIRT1 HST2, S. cerevisiae, homolog of NAD dependent deacetylase sirtuin 1 NAD dependent protein deacetylase sirtuin 1 OTTHUMP00000198111 OTTHUMP00000198112 Regulatory protein SIR2 homolog 1 SIR1_HUMAN SIR2 SIR2 like 1 SIR2 like protein 1 SIR2, S.cerevisiae, homolog-like 1 SIR2-like protein 1 SIR2ALPHA SIR2L1 Sirt1 SirtT1 75 kDa fragment Sirtuin (silent mating type information regulation 2 homolog) 1 (S. cerevisiae) Sirtuin 1 Sirtuin type 1

Images

Fig1: Western blot analysis of SIRT1 on different lysates with Rabbit anti-SIRT1 antibody (ER130811) at 1/500 dilution.

Lane 1: Hela cell lysate
Lane 2: Jurkat cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 82 kDa
Observed band size: 110 kDa

Exposure time: 2 minutes;

6% 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 (ER130811) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of SIRT1 on F9 cell lysates with Rabbit anti-SIRT1 antibody (ER130811) at 1/2,000 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 82 kDa
Observed band size: 110 kDa

Exposure time: 1 minute;

8% 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 (ER130811) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of HeLa cells labeling SIRT1 with Rabbit anti-SIRT1 antibody (ER130811) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 (ER130811) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig4: Immunocytochemistry analysis of F9 cells labeling SIRT1 with Rabbit anti-SIRT1 antibody (ER130811) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 (ER130811) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig5: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-SIRT1 antibody. Counter stained with hematoxylin.

	Fig6: Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti-SIRT1 antibody. Counter stained with hematoxylin.
	Fig7: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-SIRT1 antibody (ER130811) at 1/200 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 (ER130811) at 1/200 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.
	Fig8: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-SIRT1 antibody (ER130811) at 1/200 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 (ER130811) at 1/200 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.
	Fig9: Flow cytometric analysis of F9 cells labeling SIRT1. Cells were fixed and permeabilized. Then stained with the primary antibody (ER130811, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Fig10: Western blot analysis of SIRT1 on different lysates with Rabbit anti-SIRT1 antibody (ER130811) at 1/20,000 dilution.

Lane 1: HEK-293-si NT cell lysate
Lane 2: HEK-293-si SIRT1 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 82 kDa
Observed band size: 110 kDa

Exposure time: 50 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 (ER130811) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.