SIRT7 Recombinant Rabbit Monoclonal Antibody [PSH09-17] - BSA and Azide free

Western blot analysis of SIRT7 on different lysates with Rabbit anti-SIRT7 antibody (HA751271) at 1/2,000 dilution.

Lane 1: HEK-293 cell lysate
Lane 2: HeLa cell lysate
Lane 3: U-2 OS cell lysate
Lane 4: Neuro-2a cell lysate
Lane 5: C6 cell lysate
Lane 6: Mouse liver tissue lysate
Lane 7: Rat spleen tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 45 kDa
Observed band size: 45 kDa

Exposure time: 3 minutes; ECL: K1801;

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 (HA751271) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of RAW264.7 cells labeling SIRT7 with Rabbit anti-SIRT7 antibody (HA751271) 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-SIRT7 antibody (HA751271) 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 (HA601187, 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.

Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-SIRT7 antibody (HA751271) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (HA751271) at 1/500 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.

SIRT7 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA751271 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751271 at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA751271 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA751271 in HeLa cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 30 seconds; ECL: K1802

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with SIRT7 (HA751271) 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.