PGP9.5 Mouse Monoclonal Antibody [B1-5-6]

☑ Relative expression (RE)

Western blot analysis of PGP9.5 on different lysates with Mouse anti-PGP9.5 antibody (EM1701-86) at 1/2,000 dilution.

Lane 1: A-172 cell lysate (20 µg/Lane)
Lane 2: SHG-44 cell lysate (20 µg/Lane)
Lane 3: U-87 MG cell lysate (20 µg/Lane)
Lane 4: SH-SY5Y cell lysate (20 µg/Lane)
Lane 5: NCI-H1299 cell lysate (20 µg/Lane)
Lane 6: A549 cell lysate (20 µg/Lane)
Lane 7: 293T cell lysate (20 µg/Lane)
Lane 8: LNCaP cell lysate (negative) (20 µg/Lane)
Lane 9: K-562 cell lysate (negative) (20 µg/Lane)
Lane 10: Neuro-2a cell lysate (20 µg/Lane)
Lane 11: C6 cell lysate (20 µg/Lane)
Lane 12: PC-12 cell lysate (20 µg/Lane)
Lane 13: Mouse brain tissue lysate (20 µg/Lane)
Lane 14: Rat brain tissue lysate (20 µg/Lane)

Predicted band size: 25 kDa
Observed band size: 25 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 (EM1701-86) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Knockdown (KD)

Western blot analysis of PGP9.5 on different lysates with Mouse anti-PGP9.5 antibody (EM1701-86) at 1/2,000 dilution.

Lane 1: A549-si NT cell lysate (10 µg/Lane)
Lane 2: A549-si PGP9.5 cell lysate (10 µg/Lane)

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

Exposure time: 30 seconds; ECL: K1802;
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 (EM1701-86) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of Neuro-2a cells labeling PGP9.5 with Mouse anti-PGP9.5 antibody (EM1701-86) 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 Mouse anti-PGP9.5 antibody (EM1701-86) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.

Immunocytochemistry analysis of PC-12 cells labeling PGP9.5 with Mouse anti-PGP9.5 antibody (EM1701-86) 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 Mouse anti-PGP9.5 antibody (EM1701-86) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.

Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.

Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.

Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.

Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.

Flow cytometric analysis of Neuro-2a cells labeling PGP9.5.

Cells were fixed and permeabilized. Then stained with the primary antibody (EM1701-86, 1μg/mL) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Flow cytometric analysis of PC-12 cells labeling PGP9.5.

Cells were fixed and permeabilized. Then stained with the primary antibody (EM1701-86, 1μg/mL) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).