ATPase Inhibitory Factor 1 / IF1 Recombinant Rabbit Monoclonal Antibody [PSH16-94] - BSA and Azide free

This data was developed using HA723890, the same antibody clone in a different buffer formulation.
Western blot analysis of ATPase Inhibitory Factor 1 / IF1 on different lysates with Rabbit anti-ATPase Inhibitory Factor 1 / IF1 antibody (HA723890) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: K-562 cell lysate
Lane 3: MCF7 cell lysate
Lane 4: HepG2 cell lysate
Lane 5: HT-29 cell lysate
Lane 6: COS-1 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 12 kDa
Observed band size: 12 kDa

Exposure time: 1 minute 50 seconds; 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 (HA723890) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

This data was developed using HA723890, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HeLa cells labeling ATPase Inhibitory Factor 1 / IF1 with Rabbit anti-ATPase Inhibitory Factor 1 / IF1 antibody (HA723890) at 1/1,000 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-ATPase Inhibitory Factor 1 / IF1 antibody (HA723890) at 1/1,000 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.

This data was developed using HA723890, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-ATPase Inhibitory Factor 1 / IF1 antibody (HA723890) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA723890) at 1/5,000 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.

This data was developed using HA723890, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-ATPase Inhibitory Factor 1 / IF1 antibody (HA723890) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA723890) at 1/5,000 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.

This data was developed using HA723890, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human heart tissue with Rabbit anti-ATPase Inhibitory Factor 1 / IF1 antibody (HA723890) at 1/20,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA723890) at 1/20,000 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.

This data was developed using HA723890, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of HeLa cells labeling ATPase Inhibitory Factor 1 / IF1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA723890, 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).

This data was developed using HA723890, the same antibody clone in a different buffer formulation.
ATPase Inhibitory Factor 1 / IF1 was immunoprecipitated from 0.2 mg K-562 cell lysate with HA723890 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723890 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: K-562 cell lysate (input)
Lane 2: HA723890 IP in K-562 cell lysate
Lane 3: Rabbit IgG instead of HA723890 in K-562 cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 59 seconds; ECL: K1801