Cdk4 Recombinant Rabbit Monoclonal Antibody [SD205-1]
cat.: ET1612-23
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality: Monoclonal
Clone number: SD205-1
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 34 kDa
Isotype: IgG
Immunogen: Recombinant protein within mouse Cdk4 aa 180-303.
Positive control: SK-OV-3 cell lysate, Jurkat cell lysate, HeLa cell lysate, MCF7 cell lysate, K-562 cell lysate, A549 cell lysate, NIH:OVCAR-3 cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, C6 cell lysate, PC-12 cell lysate, Mouse brain tissue lysate, Rat lung tissue lysate, human thyroid carcinoma tissue, mouse colon tissue, mouse lung tissue, rat colon tissue, rat lung tissue, NIH/3T3, HeLa.
Subcellular location: Cytoplasm, Nucleus, Membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  IP
  FC
1:2,000
1:50-1:200
1:50-1:200
1:200-1:1,000
Use at an assay dependent concentration.
1:1,000
Uniprot #: SwissProt: P11802 Human | P30285 Mouse | P35426 Rat
Alternative names: Cdk 4 cdk4 CDK4 protein CDK4_HUMAN Cell division kinase 4 Cell division protein kinase 4 CMM 3 CMM3 Crk3 Cyclin dependent kinase 4 Cyclin-dependent kinase 4 Melanoma cutaneous malignant 3 MGC14458 p34 cdk4 PSK J3 PSK-J3
Images
ET1612-23_1.jpg Fig1: Western blot analysis of Cdk4 on different lysates with Rabbit anti-Cdk4 antibody (ET1612-23) at 1/2,000 dilution.

Lane 1: SK-OV-3 cell lysate (20 µg/Lane)
Lane 2: Jurkat cell lysate (20 µg/Lane)
Lane 3: HeLa cell lysate (20 µg/Lane)
Lane 4: MCF7 cell lysate (20 µg/Lane)
Lane 5: K-562 cell lysate (20 µg/Lane)
Lane 6: A549 cell lysate (20 µg/Lane)
Lane 7: NIH:OVCAR-3 cell lysate (20 µg/Lane)
Lane 8: NIH/3T3 cell lysate (20 µg/Lane)
Lane 9: C2C12 cell lysate (20 µg/Lane)
Lane 10: C6 cell lysate (20 µg/Lane)
Lane 11: PC-12 cell lysate (20 µg/Lane)
Lane 12: Mouse brain tissue lysate (40 µg/Lane)
Lane 13: Rat lung tissue lysate (40 µg/Lane)

Predicted band size: 34 kDa
Observed band size: 30 kDa

Exposure time: 24 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 (ET1612-23) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1612-23_2.jpg Fig2: ICC staining of Cdk4 in AGS cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-23, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1612-23_3.jpg Fig3: ICC staining of Cdk4 in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-23, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1612-23_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human thyroid carcinoma tissue with Rabbit anti-Cdk4 antibody (ET1612-23) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-23) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1612-23_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Cdk4 antibody (ET1612-23) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-23) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1612-23_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-Cdk4 antibody (ET1612-23) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-23) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1612-23_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-Cdk4 antibody (ET1612-23) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-23) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1612-23_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-Cdk4 antibody (ET1612-23) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-23) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1612-23_9.jpg Fig9: Immunocytochemistry analysis of NIH/3T3 cells labeling Cdk4 with Rabbit anti-Cdk4 antibody (ET1612-23) 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 Rabbit anti-Cdk4 antibody (ET1612-23) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1612-23_10.jpg Fig10: Flow cytometric analysis of HeLa cells labeling Cdk4.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1612-23, 1μg/mL) (red) compared with Rabbit IgG 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-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.