WB
Western blot analysis of Cdk4 on different lysates with Rabbit anti-Cdk4 antibody 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; 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human thyroid carcinoma tissue with Rabbit anti-Cdk4 antibody 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 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.ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells labeling Cdk4 with Rabbit anti-Cdk4 antibody 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of SH-SY5Y cells labeling PGP9.5. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | Cdk4 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within mouse Cdk4 aa 180-303. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:2000 IHC:1:200-1:1000 ICC:1:50-1:200 FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | CDK4 |
|---|---|
| Gene Synonyms | CMM3 PSK-J3 |
| Gene Full Name | cyclin dependent kinase 4 |
| Gene Summary | The protein encoded by this gene is a member of the Ser/Thr protein kinase family. This protein is highly similar to the gene products of S. cerevisiae cdc28 and S. pombe cdc2. It is a catalytic subunit of the protein kinase complex that is important for cell cycle G1 phase progression. The activity of this kinase is restricted to the G1-S phase, which is controlled by the regulatory subunits D-type cyclins and CDK inhibitor p16(INK4a). This kinase was shown to be responsible for the phosphorylation of retinoblastoma gene product (Rb). Mutations in this gene as well as in its related proteins including D-type cyclins, p16(INK4a) and Rb were all found to be associated with tumorigenesis of a variety of cancers. Multiple polyadenylation sites of this gene have been reported. [provided by RefSeq, Jul 2008] |
| Alternative Names | Epididymis luminal protein 117 antibody Epididymis secretory protein Li 53 antibody HEL 117 antibody HEL S 53 antibody NDGOA antibody Neuron cytoplasmic protein 9.5 antibody OTTHUMP00000218137 antibody OTTHUMP00000218139 antibody OTTHUMP00000218140 antibody OTTHUMP00000218141 antibody Epididymis luminal protein 117 antibody Epididymis secretory protein Li 53 antibody HEL 117 antibody HEL S 53 antibody NDGOA antibody Neuron cytoplasmic protein 9.5 antibody OTTHUMP00000218137 antibody OTTHUMP00000218139 antibody OTTHUMP00000218140 antibody OTTHUMP00000218141 antibody Park 5 antibody PARK5 antibody PGP 9.5 antibody PGP9.5 antibody PGP95 antibody Protein gene product 9.5 antibody Ubiquitin C terminal esterase L1 antibody Ubiquitin C terminal hydrolase antibody Ubiquitin C terminal hydrolase L1 antibody Ubiquitin carboxyl terminal esterase L1 antibody Ubiquitin carboxyl terminal hydrolase isozyme L1 antibody Ubiquitin carboxyl-terminal hydrolase isozyme L1 antibody Ubiquitin thioesterase L1 antibody Ubiquitin thiolesterase antibody Ubiquitin thiolesterase L1 antibody UCH-L1 antibody UCHL1 antibody UCHL1_HUMAN antibody |
| Molecular Weight(MW) | 34kDa(Observed band size: 30kDa) |
| Cellular Localization | Cytoplasm, Nucleus, Membrane. |
WB
Western blot analysis of Cdk4 on different lysates with Rabbit anti-Cdk4 antibody 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; 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human thyroid carcinoma tissue with Rabbit anti-Cdk4 antibody 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 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.
ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells labeling Cdk4 with Rabbit anti-Cdk4 antibody 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of SH-SY5Y cells labeling PGP9.5. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:2000 IHC:1:200-1:1000 ICC:1:50-1:200 FC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | 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. |
