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Phospho-Cdc6 (S54) Recombinant Rabbit Monoclonal Antibody [SD08-47]

Usd: 385

Specification

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Catalog# ET1612-96

Phospho-Cdc6 (S54) Recombinant Rabbit Monoclonal Antibody [SD08-47]

Application

  • WB

  • IF-Tissue

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

Phospho-Cdc6 (S54) Recombinant Rabbit Monoclonal Antibody [SD08-47]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser54 of Human Cdc6 aa 41-90 / 560.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Tissue, IHC-P

Molecular Weight

Predicted band size: 63 kDa

Positive Control

HeLa whole cell lysate, HeLa treated with 3mM Hydroxyurea for 20 hours whole cell lysate, Jurkat cell lysates, mouse spleen tissue, rat spleen tissue.

Conjugation

unconjugated

Clone Number

SD08-47

RRID

AB_3073461

Product Features

Form

Liquid

Concentration

Storage Instructions

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

Storage Buffer

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:2,000

Target

Function

Cdc6, or cell division cycle 6, is a protein in eukaryotic cells. CDC6 is an ATP binding protein and a member of the pre-replicative complex (pre-RC) together with the origin recognition complex (ORC), Cdt1 and the MCM complex (containing MCM2-7p). CDC6 assembles after ORC in an ATP dependent manner and is required for loading MCM proteins onto the DNA. It is thought that the CDC6-Cdt1 complex uses ATP hydrolysis to thread DNA through the central hole of the MCM doughnut. Mutations in the binding motif of CDC6 strongly suggest that ATP binding and hydrolysis is essential for its function. The minimal requirement for DNA binding has been mapped within its 47-amino acid sequence. Furthermore, Cdc6 indirectly inhibits activation of the p34cdc2/CDC28 M phase kinase, thus nuclear division is suppressed.

Background References

1. Fan X et al. Role of Cdc6 in re-replication in cells expressing human papillomavirus E7 oncogene. Carcinogenesis 37:799-809 (2016).

2. Huang J et al. miR-34a Modulates Angiotensin II-Induced Myocardial Hypertrophy by Direct Inhibition of ATG9A Expression and Autophagic Activity. PLoS One 9:e94382 (2014).

Sequence Similarity

Belongs to the CDC6/cdc18 family.

Subcellular Location

Nucleus, Cytoplasm.

UNIPROT

SwissProt: Q99741 Human

SwissProt: O89033 Mouse

Entrez Gene: 360621 Rat

Synonyms

Cdc 18L antibody

Cdc 6 antibody

CDC18 (cell division cycle 18, S.pombe, homolog) like antibody

CDC18 (S.pombe) antibody

Cdc18 related protein antibody

CDC18(S.pombe) antibody

Cdc18-related protein antibody

Cdc18L antibody

cdc6 antibody

CDC6 cell division cycle 6 homolog antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-Cdc6 (S54) on different lysates with Rabbit anti-Phospho-Cdc6 (S54) antibody (<a href="/products/ET1612-96" style="font-weight: bold;text-decoration: underline;">ET1612-96</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa whole cell lysate<br />Lane 2: HeLa treated with 3mM Hydroxyurea for 20 hours whole cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 63 kDa<br />Observed band size: 63 kDa<br /><br />Exposure time: 1 minute 30 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1612-96" style="font-weight: bold;text-decoration: underline;">ET1612-96</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-Cdc6 (S54) on different lysates with Rabbit anti-Phospho-Cdc6 (S54) antibody (ET1612-96) at 1/1,000 dilution.

    Lane 1: HeLa whole cell lysate
    Lane 2: HeLa treated with 3mM Hydroxyurea for 20 hours whole cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 63 kDa
    Observed band size: 63 kDa

    Exposure time: 1 minute 30 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 (ET1612-96) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Phospho-Cdc6 (S54) on Jurkat cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/ET1612-96" style="font-weight: bold;text-decoration: underline;">ET1612-96</a>, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:200,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Phospho-Cdc6 (S54) on Jurkat cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1612-96, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-Phospho-Cdc6 (S54) antibody (<a href="/products/ET1612-96" style="font-weight: bold;text-decoration: underline;">ET1612-96</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-96" style="font-weight: bold;text-decoration: underline;">ET1612-96</a>) at 1/2,000 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.

    Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-Phospho-Cdc6 (S54) antibody (ET1612-96) at 1/2,000 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 (ET1612-96) at 1/2,000 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.

  • Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-Phospho-Cdc6 (S54) antibody (<a href="/products/ET1612-96" style="font-weight: bold;text-decoration: underline;">ET1612-96</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-96" style="font-weight: bold;text-decoration: underline;">ET1612-96</a>) at 1/2,000 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.

    Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-Phospho-Cdc6 (S54) antibody (ET1612-96) at 1/2,000 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 (ET1612-96) at 1/2,000 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.

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

Citation