Recombinant Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008)

Product name

Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546]
Description

Rabbit monoclonal [EPR19546] to CDK1 (phospho T161) + CDK2 (phospho T160) + CDK3 (phospho T160)
Host species

Rabbit
Specificity

ab201008 also recognizes CDK2 (phospho T160) and CDK3 (phospho T160).
Tested applications

Suitable for: WB, Dot blot, IHC-P, IP, ICC/IFmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa treated with UV for 90 minutes, HeLa treated with UV for 90 minutes then with alkaline phosphatase for 1 hour, C6, C6 treated with alkaline phosphatase for 1 hour and NIH/3T3 whole cell lysate. IHC-P: Human tonsil and cervix cancer tissues; Rat testis tissue. ICC/IF: HeLa cells. IP: HeLa treated with 25J/m2 UV for 1-hour whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19546
Isotype

IgG
Research areas

Alternative Versions
- Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] - BSA and Azide free (ab251327)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (ab210849)
Recombinant Protein
- Recombinant Human CDK1 protein (ab187447)
Related Buffer
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)
Related Products
- Anti-CDK1 antibody [EPR165] (ab133327)
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab201008 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB		1/1000. Detects a band of approximately 34, 28 kDa (predicted molecular weight: 34 kDa).
Dot blot		1/1000.
IHC-P		1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Recommended for human and rat only.
IP		1/30.
ICC/IF		1/100.

Notes
WB 1/1000. Detects a band of approximately 34, 28 kDa (predicted molecular weight: 34 kDa).
Dot blot 1/1000.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Recommended for human and rat only.
IP 1/30.
ICC/IF 1/100.

Cellular localization

CDK1: Nucleus.
Database links
- Entrez Gene: 1017 Human
- Entrez Gene: 1018 Human
- Entrez Gene: 983 Human
- Entrez Gene: 12534 Mouse
- Entrez Gene: 12566 Mouse
- Entrez Gene: 362817 Rat
- Entrez Gene: 54237 Rat
- Omim: 116940 Human
- Omim: 116953 Human
- Omim: 123828 Human
- SwissProt: P06493 Human
- SwissProt: P24941 Human
- SwissProt: Q00526 Human
- SwissProt: P11440 Mouse
- SwissProt: P97377 Mouse
- SwissProt: Q80YP0 Mouse
- SwissProt: P39951 Rat
- SwissProt: Q63699 Rat
- Unigene: 19192 Human
- Unigene: 334562 Human
- Unigene: 689624 Human
- Unigene: 706766 Human
- Unigene: 111326 Mouse
- Unigene: 281367 Mouse
- Unigene: 33677 Mouse
- Unigene: 104460 Rat
- Unigene: 6934 Rat
see all
Form

CDK1: CDK1 can be located to the Nucleus, cytoplasm and Mithocondria. It's cytoplasmic during interphase and reversibly translocated from cytoplasm to the nucleus when phosphorilated before G2-M transition when associated with cyclin-B1. Accumulates in mitochondria in G2-arrested cells upon DNA-damage.

Western blot - Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008)

All lanes : Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008) at 1/1000 dilution

Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa treated with UV for 90 minutes whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 34 kDa
Observed band size: 28,34 kDa why is the actual band size different from the predicted?

Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Dot Blot - Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008)

Dot blot analysis of CDK1 (phospho T161) labeled with ab201008 at 1/1000 dilution.

Lane 1: CDK1 (phospho T161) phospho peptide.
Lane 2: CDK1 non-phospho peptide.

Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Dot Blot - Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008)

Dot blot analysis of CDK2 (phospho T160) labeled with ab201008 at 1/1000 dilution.

Lane 1: CDK2 (phospho T160) phospho peptide.
Lane 2: CDK2 non-phospho peptide.

Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Dot Blot - Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008)

Dot blot analysis of CDK3 (phospho T160) labeled with ab201008 at 1/1000 dilution.

Lane 1: CDK3 (phospho T160) phospho peptide.
Lane 2: CDK3 non-phospho peptide.

Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Western blot - Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008)

All lanes : Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008) at 1/1000 dilution

Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa treated with UV for 90 minutes whole cell lysate
Lane 3 : HeLa treated with UV for 90 minutes then with alkaline phosphatase for 1-hour whole cell lysate
Lane 4 : HeLa treated with UV for 90 minutes then with alkaline phosphatase overnight whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 34 kDa
Observed band size: 28,34 kDa why is the actual band size different from the predicted?

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008)

All lanes : Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008) at 1/1000 dilution

Lane 1 : C6 (rat glial tumor cell line) whole cell lysate
Lane 2 : C6 treated with alkaline phosphatase for 1 hour whole cell lysate
Lane 3 : C6 treated with alkaline phosphatase overnight whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 34 kDa
Observed band size: 28,34 kDa why is the actual band size different from the predicted?

Exposure time: 10 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008)

All lanes : Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008) at 1/1000 dilution

Lane 1 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
Lane 2 : C6 (rat glial tumor cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 34 kDa
Observed band size: 28 kDa why is the actual band size different from the predicted?

Exposure time: 3 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CDK1 (phospho T161) with ab201008 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus and weak cytoplasm staining of germinal center from human tonsil is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008)

Immunohistochemical analysis of paraffin-embedded human cervix cancer tissue labeling CDK1 (phospho T161) with ab201008 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus and cytoplasm staining of cancer cells from human cervix cancer is observed [PMID: 15623629]. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008)

Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling CDK1 (phospho T161) with ab201008 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus and cytoplasm staining of rat testis is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling CDK1 (phospho T161) with ab201008 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing specific signal in M phase cells. The signal decreased after treatment with lambda protein phosphatase 31°C for 5 hours.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-Alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
Immunoprecipitation - Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008)

CDK1 (phospho T161) was immunoprecipitated from 0.35mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab201008 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab201008 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: HeLa treated with 25J/m2 UV for 1 hour whole cell lysate 10 µg (Input).
Lane 2: ab201008 IP in HeLa treated with 25J/m2 UV for 1 hour whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab201008 in HeLa treated with 25J/m2 UV for 1 hour whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 minutes.
Anti-CDK1 (phospho T161) + CDK2 / CDK3 (phospho T160) antibody [EPR19546] (ab201008)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab201008? Please let us know so that we can cite the reference in this datasheet.

ab201008 has been referenced in 7 publications.

Zhang Y et al. LncRNA-CCDC144NL-AS1 Promotes the Development of Hepatocellular Carcinoma by Inducing WDR5 Expression via Sponging miR-940. J Hepatocell Carcinoma 8:333-348 (2021). PubMed: 33977095
Hu S et al. LINCS gene expression signature analysis revealed bosutinib as a radiosensitizer of breast cancer cells by targeting eIF4G1. Int J Mol Med 47:N/A (2021). PubMed: 33693953
Zhao J et al. lncRNA LIFR-AS1 inhibits gastric carcinoma cell proliferation, migration and invasion by sponging miR-4698. Mol Med Rep 23:N/A (2021). PubMed: 33355363
Yang T et al. Punicalin Alleviates OGD/R-Triggered Cell Injury via TGF-ß-Mediated Oxidative Stress and Cell Cycle in Neuroblastoma Cells SH-SY5Y. Evid Based Complement Alternat Med 2021:6671282 (2021). PubMed: 33628309
Huang W et al. Blocking the short isoform of augmenter of liver regeneration inhibits proliferation of human multiple myeloma U266 cells via the MAPK/STAT3/cell cycle signaling pathway. Oncol Lett 21:197 (2021). PubMed: 33574936
Chen X et al. Incaspitolide A extracted from Carpesium cernuum induces apoptosis in vitro via the PI3K/AKT pathway in benign prostatic hyperplasia. Biosci Rep 41:N/A (2021). PubMed: 34100062
He P et al. AMPK Activity Contributes to G2 Arrest and DNA Damage Decrease via p53/p21 Pathways in Oxidatively Damaged Mouse Zygotes. Front Cell Dev Biol 8:539485 (2020). PubMed: 33015052

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Buffer

Related Products

Applications

The Abpromise guarantee

Target

Cellular localization

Database links

Form

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (7)

Customer reviews and Q&As