- PA5-27827 - Invitrogen Antibodies CDK4 antibody

Hu X, Lou T, Yuan C, Wang Y, Tu X, Wang Y, Zhang T
Oncology letters 2021 Aug;22(2):621

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of CDK4 using 30 µg of 293T lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a CDK4 polyclonal antibody (Product # PA5-27827) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of CDK4 using 30 µg of NIH-3T3 lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a CDK4 polyclonal antibody (Product # PA5-27827) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of A-431 (Lane 1), U-87 MG (Lane 2), K-562 (Lane 3), HeLa (Lane 4) and A549 (Lane 5). The blot was probed with Anti-CDK4 Polyclonal Antibody (Product # PA5-27827, 1:500 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 ~35 kDa band corresponding to CDK4 was observed across the cell lines tested. A non-specific band was also observed at around 55 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of CDK4 was performed by separating 30 µg of various whole cell extracts by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a CDK4 Polyclonal Antibody (Product # PA5-27827) at a dilution of 1:1000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using CDK4 Polyclonal Antibody (Product # PA5-27827). Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with CDK4 Polyclonal Antibody (Product # PA5-27827) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of CDK4 was performed by separating 30 µg of whole cell extract by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a CDK4 Polyclonal Antibody (Product # PA5-27827) at a dilution of 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using CDK4 Polyclonal Antibody (Product # PA5-27827). Whole cell extract (30 µg) was separated by 12% SDS-PAGE, and the membrane was blotted with CDK4 Polyclonal Antibody (Product # PA5-27827) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of CDK4 was achieved by transfecting A-431 cells with CDK4 specific validated siRNA (Silencer® select Product # s2823). Western blot analysis (Fig. a) was performed using whole cell extracts from the CDK4 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-CDK4 Polyclonal Antibody (Product # PA5-27827, 1:500 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). 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 CDK4.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of CDK4 was performed in MCF7 cells fixed in 4% paraformaldehyde at RT for 15 min. Green: CDK4 Polyclonal Antibody (Product # PA5-27827) diluted at 1:1000. Red: p21 Cip1, a nucleus marker. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of CDK4 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: CDK4 Polyclonal Antibody (Product # PA5-27827) diluted at 1:500. Red: phalloidin, a cytoskeleton marker. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of CDK4 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 CDK4 Rabbit Polyclonal Antibody(Product # PA5-27827) at 5 µg/mL 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 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of paraffin-embedded human gastric cancer, using CDK4 (Product # PA5-27827) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6. Cell cycle distribution of HGC-27 and drug-resistant cell lines. Cell cycle distribution of (A) HGC-27 and (B) HGC-27 DDP-resistant cell lines was analyzed using flow cytometry. Expression of proteins related to the cell cycle in (C) HGC-27 and (D) HGC-27 DDP-resistant cell lines was analyzed using western blotting. # P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 7. Cell cycle distribution of FU97 and drug-resistant cell lines. Cell cycle distribution of (A) FU97 and (B) FU97 DDP-resistant cell lines was detected using flow cytometry. Expression of proteins related to the cell cycle in (C) FU97 and (D) FU97 DDP-resistant cell lines was analyzed using western blotting. ## P

PA5-27827