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CDk1 Rabbit Polyclonal Antibody

Usd: 315

Specification

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Catalog# ER31213

CDk1 Rabbit Polyclonal Antibody

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

CDk1 Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Synthetic peptide within N-terminal human CDk1.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC

Target Molecular Weight

Predicted band size: 34 kDa

Positive Control

MCF-7 cell lysate, Jurkat cell lysate, PC12 cell lysate, HepG2 cell lysate, Hela cell lysate, NIH/3T3 cell lysate, mouse liver tissue lysate, SKBR3 cell lysate, HeLa, NIH/3T3, rat spleen tissue, human tonsil tissue, human breast cancer tissue, mouse spleen tissue.

Conjugation

unconjugated

RRID

AB_3069564

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:500

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:200

  • FC

  • 1:1,000

Target

Function

Cdk1 is a small protein (approximately 34 kilodaltons), and is highly conserved. Cdk1 is comprised mostly by the bare protein kinase motif, which other protein kinases share. Cdk1, like other kinases, contains a cleft in which ATP fits. When bound to its cyclin partners, Cdk1 phosphorylation leads to cell cycle progression. Given its essential role in cell cycle progression, Cdk1 is highly regulated. Most obviously, Cdk1 is regulated by its binding with its cyclin partners. Cyclin binding alters access to the active site of Cdk1, allowing for Cdk1 activity; furthermore, cyclins impart specificity to Cdk1 activity. At least some cyclins contain a hydrophobic patch which may directly interact with substrates, conferring target specificity. Furthermore, cyclins can target Cdk1 to particular subcellular locations.

Background References

1. "Sequential phosphorylation of Nedd1 by Cdk1 and Plk1 is required for targeting of the gammaTuRC to the centrosome." Zhang X., Chen Q., Feng J., Hou J., Yang F., Liu J., Jiang Q., Zhang C. J. Cell Sci. 122:2240-2251(2009)

2. "Cdc25 phosphatases are required for timely assembly of CDK1-cyclin B at the G2/M transition." Timofeev O., Cizmecioglu O., Settele F., Kempf T., Hoffmann I. J. Biol. Chem. 285:16978-16990(2010)

Sequence Similarity

Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.

Tissue Specificity

Isoform 2 is found in breast cancer tissues.

Post-translational Modification

Phosphorylation at Thr-161 by CAK/CDK7 activates kinase activity. Phosphorylation at Thr-14 and Tyr-15 by PKMYT1 prevents nuclear translocation. Phosphorylation at Tyr-15 by WEE1 and WEE2 inhibits the protein kinase activity and acts as a negative regulator of entry into mitosis (G2 to M transition). Phosphorylation by PKMYT1 and WEE1 takes place during mitosis to keep CDK1-cyclin-B complexes inactive until the end of G2. By the end of G2, PKMYT1 and WEE1 are inactivated, but CDC25A and CDC25B are activated. Dephosphorylation by active CDC25A and CDC25B at Thr-14 and Tyr-15, leads to CDK1 activation at the G2-M transition. Phosphorylation at Tyr-15 by WEE2 during oogenesis is required to maintain meiotic arrest in oocytes during the germinal vesicle (GV) stage, a long period of quiescence at dictyate prophase I, leading to prevent meiotic reentry. Phosphorylation by WEE2 is also required for metaphase II exit during egg activation to ensure exit from meiosis in oocytes and promote pronuclear formation. Phosphorylated at Tyr-4 by PKR/EIF2AK2 upon genotoxic stress. This phosphorylation triggers CDK1 polyubiquitination and subsequent proteolysis, thus leading to G2 arrest. In response to UV irradiation, phosphorylation at Tyr-15 by PRKCD activates the G2/M DNA damage checkpoint.; Polyubiquitinated upon genotoxic stress.

Subcellular Location

Cytoplasm, nucleus, Cytoskeleton, Mitochondrion.

UNIPROT

SwissProt: P06493 Human

Synonyms

Cdc 2 antibody

Cdc2 antibody

CDC28A antibody

CDK 1 antibody

CDK1 antibody

CDK1_HUMAN antibody

CDKN1 antibody

CELL CYCLE CONTROLLER CDC2 antibody

Cell division control protein 2 antibody

Cell division control protein 2 homolog antibody

Expand

Images

  • Immunocytochemistry analysis of HeLa cells labeling CDk1 with Rabbit anti-CDk1 antibody (<a href="/products/ER31213" style="font-weight: bold;text-decoration: underline;">ER31213</a>) at 1/100 dilution.<br /><br />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-CDk1 antibody (<a href="/products/ER31213" style="font-weight: bold;text-decoration: underline;">ER31213</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling CDk1 with Rabbit anti-CDk1 antibody (ER31213) 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-CDk1 antibody (ER31213) 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.

  • Immunocytochemistry analysis of NIH/3T3 cells labeling CDk1 with Rabbit anti-CDk1 antibody (<a href="/products/ER31213" style="font-weight: bold;text-decoration: underline;">ER31213</a>) at 1/100 dilution.<br /><br />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-CDk1 antibody (<a href="/products/ER31213" style="font-weight: bold;text-decoration: underline;">ER31213</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of NIH/3T3 cells labeling CDk1 with Rabbit anti-CDk1 antibody (ER31213) 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-CDk1 antibody (ER31213) 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.

  • Western blot analysis of CDk1 on different lysates using anti-CDk1 antibody at 1/500 dilution.<br /><span style="font-weight: bold;">Positive control:</span> <br />Lane 1: MCF-7 cell lysate <br />Lane 2: Jurkat cell lysate <br />Lane 3: PC12 cell lysate <br />Lane 4: HepG2 cell lysate <br />Lane 5: Hela cell lysate <br />Lane 6: NIH/3T3 cell lysate <br />Lane 7: Mouse liver tissue lysate <br />Lane 8: SKBR3 cell lysate

    Western blot analysis of CDk1 on different lysates using anti-CDk1 antibody at 1/500 dilution.
    Positive control:
    Lane 1: MCF-7 cell lysate
    Lane 2: Jurkat cell lysate
    Lane 3: PC12 cell lysate
    Lane 4: HepG2 cell lysate
    Lane 5: Hela cell lysate
    Lane 6: NIH/3T3 cell lysate
    Lane 7: Mouse liver tissue lysate
    Lane 8: SKBR3 cell lysate

  • Immunohistochemical analysis of paraffin-embedded rat spleen tissue using anti-CDk1 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded rat spleen tissue using anti-CDk1 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CDk1 antibody. Counter stained with hematoxylin

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CDk1 antibody. Counter stained with hematoxylin

  • Immunohistochemical analysis of paraffin-e.mbedded human breast cancer tissue using anti-CDk1 antibody. Counter stained with hematoxylin

    Immunohistochemical analysis of paraffin-e.mbedded human breast cancer tissue using anti-CDk1 antibody. Counter stained with hematoxylin

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-CDk1 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-CDk1 antibody. Counter stained with hematoxylin.

  • Flow cytometric analysis of HeLa cells labeling CDk1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ER31213" style="font-weight: bold;text-decoration: underline;">ER31213</a>, 1/1,000) (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&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling CDk1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ER31213, 1/1,000) (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).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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