Anti-PPT1/PPT 抗体 [EPR27163-2] (ab302895)

Blocking / Diluting buffer and concentration: 5% NFDM/TBST

Exposure time:

Lane 1: 10 seconds
Lanes 2-3: 26 seconds
Lane 4: 81 seconds

The molecular weight observed is consistent with what has been described in the literature (PMID: 28878621).

Blocking / Diluting buffer and concentration: 5% NFDM/TBST

Exposure time:

Lane 1: 15 seconds
Lanes 2-3: 180 seconds

Immunohistochemical analysis of paraffin-embedded human cerebellum tissue labeling PPT1/PPT with ab302895 at 1/5000 dilution (0.092 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection). Positive staining on human cerebellum. The section was incubated with ab302895 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval solution 2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling PPT1/PPT with ab302895 at 1/5000 dilution (0.092 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection). Positive staining on human cerebrum. The section was incubated with ab302895 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval solution 2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling PPT1/PPT with ab302895 at 1/5000 dilution (0.092 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection). Positive staining on human lung cancer. The section was incubated with ab302895 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval solution 2) for 20 mins.

Immunofluorescent analysis of 80% Methanol-fixed, 0.1% TritonX-100 permeabilized THP-1 (human monocytic leukemia monocyte) cells labeling PPT1/PPT with ab302895 at 1/50 (9.24 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2µg/ml) dilution (Green). Confocal image showing cytoplasmic staining in THP-1 cell line. is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5µg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2µg/ml) dilution.

Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized THP-1 (human monocytic leukemia monocyte) cells labeling PPT1/PPT with ab302895 at 1/500 dilution (0.1 µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

PPT1/PPT was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte), whole cell lysate 10 µg with ab302895 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302895 at ab302895 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1: THP-1 (human monocytic leukemia monocyte), whole cell lysate 10 µg

Lane 2: ab302895 IP in THP-1 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab302895 in THP-1 whole cell lysate

Observed MW (kDd): 31-35

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 10 seconds