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CDK4 antibody - C-terminal region (ARP30259_P050)

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Description of Target:
Cdk4 is probably involved in the control of the cell cycle.
Gene Symbol:
CDK4
Official Gene Full Name:
Cyclin-dependent kinase 4
NCBI Gene Id:
1019
Alias Symbols:
CMM3; MGC14458; PSK-J3
Sample Type Confirmation:

CDK4 is supported by BioGPS gene expression data to be expressed in HCT116

Tissue Tool:
Find tissues and cell lines supported to express CDK4.
Protein Accession #:
NP_000066
Nucleotide Accession#:
NM_000075
Swissprot Id:
P11802
Host:
Rabbit
Clonality:
Polyclonal
Protein Name:
Cyclin-dependent kinase 4
Protein Size (# AA):
303
Molecular Weight:
34kDa
Application:
IHC, WB
Partner Proteins:
MYC,BRCA1,CCND1,CCND1,CCND2,CDKN1A,CDKN2A,CDKN2A,CDKN2A,CDKN2A,CDKN2C,RB1,SMAD2,SMAD3,tax,BAG6,BIRC5,BMPR1B,BRCA1,CAPNS1,CCND1,CCND2,CCND3,CDC37,CDC45,CDC7,CDKN1A,CDKN1B,CDKN1C,CDKN2A,CDKN2B,CDKN2C,CDKN2D,CEBPA,CNOT7,IFI27,MCM2,MYOD1,NCOA2,NOL12,ORC3,PPP2R1B,PRKAR1A,PSMD10,PTMA,RB1,RBL1,RFC1,RFC4,SERTAD1,SETDB1,SMAD2,SMAD3,TGFBR1,VTA1,ZNF655,ACTR3B,BAG6,BIRC5,BRCA1,CAPNS1,CCND1,CCND2,CCND3,CDC37,CDKN1A,CDKN1B,CDKN2A,CDKN2B,CDKN2C,CDKN2D,CDT1,CEBPA,CNOT7,DBNL,GEMIN4,HSP82,HSPA8,IME2,LYN,MYOD1,NCOA2,PCNA,PRMT5,RB1,RFC1,RPRM,SAP25,SERTAD1,SETDB1,SMC1A,SMC3,SP1,TSPYL2,UBXN1,VTA1,WDR77
Product Format:
Lyophilized powder
Purification:
Affinity Purified
Complete computational species homology data:
CDK4 antibody - C-terminal region (ARP30259_P050)
Predicted Homology Based on Immunogen Sequence:
Human: 100%; Pig: 92%; Dog: 83%; Rat: 83%; Horse: 83%; Rabbit: 83%; Guinea pig: 83%; Sheep: 75%; Bovine: 75%
Species Reactivity:
Human, Pig, Dog, Horse, Rabbit, Rat, Guinea pig, Sheep, Bovine
Datasheets / Downloads:
Printable datasheet for
anti-CDK4 antibody
- ARP30259_P050
Peptide Sequence:
Synthetic peptide located within the following region: PRPVQSVVPEMEESGAQLLLEMLTFNPHKRISAFRALQHSYLHKDEGNPE
Blocking Peptide:
For anti-CDK4 antibody is Catalog # AAP30259
Reconstitution and Storage:
Add 50 ul of distilled water. Final anti-CDK4 antibody concentration is 1 mg/ml in PBS buffer with 2% sucrose. For longer periods of storage, store at -20C. Avoid repeat freeze-thaw cycles.

Computational species homology for CDK4 antibody (ARP30259)

Product page for CDK4 antibody (ARP30259)

The information below lists the predicted species and target name associated to the peptide antigen. Please note, all available target reference numbers to the antigen sequence are presented, including unreviewed and protein isoforms.
To search for antibodies by species, please visit Aviva’s Species Reactivity Page. To search Aviva’s catalog of antibodies by sequence, please visit Aviva’s Antibody Blast Tool.

Predicted Species & Target Target Reference Predicted Homology
Dog CDK4 antibody; Canis familiaris CDK4 antibody E2QUY0 83%
Giant panda LOC100481236 antibody; Ailuropoda melanoleuca LOC100481236 antibody D2H394 83%
Guinea pig LOC100717960 antibody; Cavia porcellus LOC100717960 antibody H0UZ32 83%
Horse LOC100051005 antibody; Equus caballus LOC100051005 antibody F6QM47 83%
Human CDK4 antibody; Homo sapiens CDK4 antibody P11802 100%
Human CDK4 antibody; Homo sapiens CDK4 antibody Q6LC83 100%
Human CDK4 antibody; Homo sapiens CDK4 antibody F8VZ13 100%
Human CDK4 antibody; Homo sapiens CDK4 antibody B4DNF9 100%
Lowland gorilla CDK4 antibody; Gorilla gorilla gorilla CDK4 antibody G3QGD2 100%
Northern white-cheeked gibbon LOC100594967 antibody; Nomascus leucogenys LOC100594967 antibody G1S5W7 100%
Pig CDK4 antibody; Sus scrofa CDK4 antibody P79432 91%
Rabbit CDK4 antibody; Oryctolagus cuniculus CDK4 antibody G1T3E1 83%
Rat CDK4 antibody; Rattus norvegicus CDK4 antibody P35426 75%
Rat Cdk4 antibody; Rattus norvegicus Cdk4 antibody F1LPI0 75%
Rhesus macaque CDK4 antibody; Macaca mulatta CDK4 antibody F7HCW9 100%
Small-eared galago CDK4 antibody; Otolemur garnettii CDK4 antibody H0XB36 90%
White-tufted-ear marmoset LOC100408304 antibody; Callithrix jacchus LOC100408304 antibody F7E4R8 100%
White-tufted-ear marmoset LOC100408304 antibody; Callithrix jacchus LOC100408304 antibody F7DPY6 100%

Product Review: CDK4 antibody - C-terminal region (ARP30259_P050) in Human colon carcinoma cell line HCT116 using Western Blot

Product Page for CDK4 antibody - C-terminal region (ARP30259_P050)

Researcher: Anke Rauch & Dr. Oliver Krämer, Institute for Biochemistry and Biophysics, Friedrich-schiller University Jena
Application: Western Blotting
Species+tissue/cell type: Human colon carcinoma cell line HCT116
How many ug’s of tissue/cell lysate run on the gel:
1. 30 ug human HCT116 cell lysate
2. 30 ug genotoxic treated human HCT116 cell lysate
3. 30 ug genotoxic treated human HCT116 cell lysate
Primary antibody dilution: 1:2000
Secondary antibody: Anti-rabbit HRP
Secondary antibody dilution: 1:5000

Questionnaire:
How do Aviva’s reagents play a role in your experimental goals? The Aviva antibody against CDK4 is used to investigate the influence of genotoxic agents on cell cycle regulation and survival. In this contest protein levels of CDK4 shall give information about the differential induction of cell cycle arrest upon genotoxic stress in human colon carcinoma cells.
How would you rate this antibody on a scale from 1-5 (5=best) and why? Antibody is rated at 5. Signals are precise, without unspecific bands.
Would you use this antibody in future experiments? Yes.
Have you used another antibody which has worked in your application? No.
Do you believe the information about the reagent on Aviva’s website is correct? The antibody was applied in a recommended concentration of 0.5 ug/ml and principly worked. Nevertheless the immunoblot presented on the datasheet showed nearly no unspecific bands.
If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not? The antibody shall be used in further experiements and data are planned to be published.
How did you store the antibody after re-suspension? Resuspended in distilled water, stored at – 20 degree C.
Sample Description, Species, Tissue/Cell Type: Human colon carcinoma cell line HCT116, 30ug protein per lane has been loaded.
How many different experimental trials were conducted using the antibody sample? Two experimental trials.
What type of experimental sample are you using and how did you preparing it? Cells were harvested from cell culture plates by scraping. Cell suspension was transferred into 1.5ml tubes and  centrifuged for 2 minutes at 700g. After discarding the supernatant cells were incubated with 150 ul lysis buffer (containing 10 % (v/v) Glycerol, 0.5% (v/v) NP-40, 0.1% (v/v) NaF, 1% (v/v) NaV and 0.2 % protease inhibitor cocktail) on ice. Subsequently cells were sonified with 30 % amplitude for 5 seconds. Samples were centrifuged for 10 minute at 14000 rpm and supernatant was used for westernblot analysis.
Primary used and dilution: Primary antibody was diluted 1:2000 and applied for 15 hours.
Secondary used and dilution: Secondary antibody against rabbit-IgG (Santa Cruz) was appliedin a 1:5000 dilution and incubated for 1 hour.
What controls were used in your experiment? Please include your positive control: Untreated cells were used as a negative control for genotoxic stress.
Experimental Procedure/Protocols: Proteins were blotted on a PVDF membrane. Thereafter the membrane was incubated in 5% milk and PBS-T (PBS + 0.05% Tween20 (Roth)) at room temperature for 1 h to block all aunspecific protein binding sites. Specific primary and secondary antibodies were used for protein detection. Primary antibodies were dilluted in 2% milk PBS-T and overnight at 4 degree C on a roll mixer. After incubation the membrane was washed 3 times with 15ml PBS-T for 10 minutes and specific secondary antibody against rabbit immunogloulins was applied for 1 h at room temperature. The membrane was washed 3 times for 10 minutes in PBS-T to rinse off the extra secondary antibodies. The membrane was transferred onto a plastic bag and incubated with SuperSignal West Pico Chemiluminescent Substrate, Thermo Scientific (1:1) for 2 minutes before it was developed.

Product Review: CDK4 antibody-C-terminal region (ARP30259_P050) in Human colon carcinoma cell line HCT116 using Western Blot

Product Page for CDK4 antibody-C-terminal region (ARP30259_P050)

Researcher: Anke Rauch & Dr. Oliver Krämer, Institute for Biochemistry and Biophysics, Center for Molecular Biomedicine, Friedrich-Schiller University Jena
Application: Western blotting
Species+tissue/cell type: Human colon carcinoma cell line HCT116
How many ug’s of tissue/cell lysate run on the gel:
1: 30 ug untreated human HCT116 cell lysate
2: 30 ug genotoxic agent treated human HCT116 cell lysate
3: 30 ug genotoxic agent treated human HCT116 cell lysate
Primary antibody dilution: 1:2000
Secondary antibody: Anti-rabbit HRP
Secondary antibody dilution: 1:5000

Questionnaire:
How do Aviva’s reagents play a role in your experimental goals? The Aviva antibody against CDK4 is used to investigate the influence of genotoxic agents on cell cycle regulation and survival. In this contest protein levels of CDK4 shall give information about the differential induction of cell cycle arrest upon genotoxic stress in human colon carcinoma cells.
How would you rate this antibody on a scale from 1-5 (5=best) and why? Antibody is rated at 5. Signals are precise, without unspecific bands.
Would you use this antibody in future experiments? Yes.
Have you used another antibody which has worked in your application? No.
Do you believe the information about the reagent on Aviva’s website is correct? The antibody was applied in a recommended concentration of 0.5 ug/ml and principly worked. Nevertheless the immunoblot presented on the datasheet showed nearly no unspecific bands.
If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not? The antibody shall be used in further experiements and data are planned to be published.
How did you store the antibody after re-suspension? Resuspended in distilled water, stored at – 20 degree C.
Sample Description, Species, Tissue/Cell Type: Human colon carcinoma cell line HCT116, 30ug protein per lane has been loaded.
How many different experimental trials were conducted using the antibody sample? Two experimental trials.
What type of experimental sample are you using and how did you preparing it? Cells were harvested from cell culture plates by scraping. Cell suspension was transferred into 1.5ml tubes and  centrifuged for 2 minutes at 700g. After discarding the supernatant cells were incubated with 150 ul lysis buffer (containing 10 % (v/v) Glycerol, 0.5% (v/v) NP-40, 0.1% (v/v) NaF, 1% (v/v) NaV and 0.2 % protease inhibitor cocktail) on ice. Subsequently cells were sonified with 30 % amplitude for 5 seconds. Samples were centrifuged for 10 minute at 14000 rpm and supernatant was used for westernblot analysis.
Primary used and dilution: Primary antibody was diluted 1:2000 and applied for 15 hours.
Secondary used and dilution: Secondary antibody against rabbit-IgG (Santa Cruz) was appliedin a 1:5000 dilution and incubated for 1 hour.
What controls were used in your experiment? Please include your positive control: Untreated cells were used as a negative control for genotoxic stress.
Experimental Procedure/Protocols: Proteins were blotted on a PVDF membrane. Thereafter the membrane was incubated in 5% milk and PBS-T (PBS + 0.05% Tween20 (Roth)) at room temperature for 1 h to block all aunspecific protein binding sites. Specific primary and secondary antibodies were used for protein detection. Primary antibodies were dilluted in 2% milk PBS-T and overnight at 4 degree C on a roll mixer. After incubation the membrane was washed 3 times with 15ml PBS-T for 10 minutes and specific secondary antibody against rabbit immunogloulins was applied for 1 h at room temperature. The membrane was washed 3 times for 10 minutes in PBS-T to rinse off the extra secondary antibodies. The membrane was transferred onto a plastic bag and incubated with SuperSignal West Pico Chemiluminescent Substrate, Thermo Scientific (1:1) for 2 minutes before it was developed.
 
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