PGP9.5 Rabbit Polyclonal Antibody

☑ Knockdown (KD)

Western blot analysis of PGP9.5 on different lysates with Rabbit anti-PGP9.5 antibody (R1511-8) at 1/1,000 dilution.

Lane 1: A549-si NT cell lysate
Lane 2: A549-si PGP9.5 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 25 kDa
Observed band size: 25 kDa

Exposure time: 3 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 (R1511-8) at 1/1,000 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.

Western blot analysis of PGP9.5 on different lysates with Rabbit anti-PGP9.5 antibody (R1511-8) at 1/2,000 dilution.

Lane 1: A549 cell lysate
Lane 2: DU 145-1 cell lysate
Lane 3: SH-SY5Y cell lysate
Lane 4: Neuro-2a cell lysate
Lane 5: C6 cell lysate

Lysates/proteins at 15 µg/Lane.
Exposure time: 8 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: R1511-8, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 25 kDa
Observed band size: 25 kDa

ICC staining of PGP9.5 in N2A cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (R1511-8, 1/200) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-PGP9.5 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (R1511-8, 1/200) for 30 minutes 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.

Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue using anti-PGP9.5 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (R1511-8, 1/200) for 30 minutes 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.

Flow cytometric analysis of PGP9.5 was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (R1511-8, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).