MA1-90008

antibody from Invitrogen Antibodies

Targeting: UCHL1 PARK5, PGP9.5, Uch-L1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-90008

Provider

Invitrogen Antibodies

Product name

PGP9.5 Monoclonal Antibody (13C4)

Antibody type

Monoclonal

Antigen

Other

Description

Recommended positive controls: Human Brain Tissue.

Antibody clone number

13C4

Concentration

0.3 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of Neuro-2a (Lane 1), SH-SY5Y (Lane 2), U-87 MG (Lane 3), PC-12 (Lane 4), HEK-293 (Lane 5), A549 (Lane 6), HeLa (Lane 7), Hep G2 (Lane 8), NIH/3T3 (Lane 9), tissue extracts of Mouse Brain (Lane 10) and Rat Brain (Lane 11). The blot was probed with Anti-PGP9.5 Monoclonal Antibody (Product # MA1-90008, 1:500 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). A 27 kDa band corresponding to PGP9.5 was detected across the cell lines and tissues tested except for Hela, HepG2 and NIH/3T3 which is reported to be negative for PGP9.5 expression.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of Neuro-2a (Lane 1), SH-SY5Y (Lane 2), U-87 MG (Lane 3), PC-12 (Lane 4), HEK-293 (Lane 5), A549 (Lane 6), HeLa (Lane 7), Hep G2 (Lane 8), NIH/3T3 (Lane 9), tissue extracts of Mouse Brain (Lane 10) and Rat Brain (Lane 11). The blot was probed with Anti-PGP9.5 Monoclonal Antibody (Product # MA1-90008, 1:500 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). A 27 kDa band corresponding to PGP9.5 was detected across the cell lines and tissues tested except for Hela, HepG2 and NIH/3T3 which is reported to be negative for PGP9.5 expression.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of PGP9.5/UCHL1 was performed by loading 20 µg of SH-SY5Y wild type (Lane 1), SH-SY5Y Cas9 control (Lane 2), SH-SY5Y PGP9.5/UCHL1 knockout (Lane 3) whole cell extracts. The blot was probed with Anti-PGP9.5/ UCHL1 Monoclonal Antibody (Product # MA1-90008) (1:500 dilution) and Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177) (1:4000 dilution). Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to PGP9.5/UCHL1.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of PGP9.5 was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PGP9.5 Monoclonal Antibody (13C4) (Product # MA1-90008) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.