- PA5-21411 - Invitrogen Antibodies PUF60 antibody

No submitted references: Submit reference

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using PUF60 Polyclonal Antibody (Product # PA5-21411). Sample (30 µg of whole cell lysate). A: Hep G2. B: MOLT4SDS PAGE. Z. C: Raji. 7.5% SDS PAGE. PUF60 Polyclonal Antibody (Product # PA5-21411) diluted at 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of PUF60 was achieved by transfecting HeLa cells with PUF60 specific siRNAs (Silencer® select Product # s223538, s22446). Western blot analysis (Fig. a) was performed using nuclear extracts from the PUF60 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with PUF60 Polyclonal Antibody (Product # PA5-21411, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to PUF60.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on nuclear enriched cell extract (30 µg lysate) of HepG2 (Lane 1), MOLT-4 (Lane 2), Raji (Lane 3), A431 (Lane 4), HeLa (Lane 5), THP-1 (Lane 6), A549 (Lane 7), and Mouse thymus (Lane 8). The blot was probed with Anti-PUF60 Polyclonal Antibody (Product # PA5-21411, 1:2000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 60 kDa band corresponding to PUF60 was detected in cell lines tested along with a non-specific band at 30 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of PUF60 in paraformaldehyde-fixed A431 cells using a PUF60 polyclonal antibody (Product # PA5-21411) at a 1:500 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of paraformaldehyde-fixed A431, using PUF60 antibody (Product # PA5-21411) at 1:500 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of PUF60 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PUF60 Polyclonal Antibody (Product # PA5-21411) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of paraffin-embedded human oral tissue, using PUF60 (Product # PA5-21411) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

PA5-21411