Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [3]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-21411 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PUF60 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, H1299, HeLaS3, HepG2, Molt-4, Raji. Predicted reactivity: Mouse (98%), Rat (98%), Bovine (98%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.81 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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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.
- 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.
- 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.
- 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.
- 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.