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Cdc25C Recombinant Rabbit Monoclonal Antibody [JE59-53]

Usd: 385

Specification

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Catalog# HA722069

Cdc25C Recombinant Rabbit Monoclonal Antibody [JE59-53]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

Conjugation

  • unconjugated

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

Overview

Product Name

Cdc25C Recombinant Rabbit Monoclonal Antibody [JE59-53]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Cdc25C aa 400-473 (C terminal).

Species Reactivity

Human

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 53 kDa

Positive Control

293T cell lysate, Caco-2 cell lysate, HUVEC cell lysate, HT-29 cell lysate, HeLa, human breast cancer tissue.

Conjugation

unconjugated

Clone Number

JE59-53

RRID

AB_3096764

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-Cell

  • 1:100

  • IHC-P

  • 1:2,000

  • FC

  • 1:1,000

Target

Function

M-phase inducer phosphatase 3 is an enzyme that in humans is encoded by the CDC25C gene. This gene is highly conserved during evolution and it plays a key role in the regulation of cell division. The encoded protein is a tyrosine phosphatase and belongs to the Cdc25 phosphatase family. It directs dephosphorylation of cyclin B-bound CDC2 (CDK1) and triggers entry into mitosis. It is also thought to suppress p53-induced growth arrest. Multiple alternatively spliced transcript variants of this gene have been described, however, the full-length nature of many of them is not known.

Background References

1. Liu K et al. The role of CDC25C in cell cycle regulation and clinical cancer therapy: a systematic review. Cancer Cell Int. 2020 Jun

2. Zhou C et al. CDC25C is a prognostic biomarker and correlated with mitochondrial homeostasis in pancreatic adenocarcinoma. Bioengineered. 2022 May

Subcellular Location

Nucleus.

UNIPROT

SwissProt: P30307 Human

Synonyms

CDC 25 antibody

Cdc 25C antibody

CDC25 antibody

CDC25C antibody

Cell division cycle 25 homolog C antibody

Cell division cycle 25C antibody

Cell division cycle 25C protein antibody

Dual specificity phosphatase Cdc25C antibody

M phase inducer phosphatase 3 antibody

M-phase inducer phosphatase 3 antibody

Expand

Images

  • Western blot analysis of Cdc25C on different lysates with Rabbit anti-Cdc25C antibody (<a href="/products/HA722069" style="font-weight: bold;text-decoration: underline;">HA722069</a>) at 1/1,000 dilution.<br /><br />Lane 1: 293T cell lysate<br />Lane 2: Caco-2 cell lysate<br />Lane 3: HUVEC cell lysate<br />Lane 4: HT-29 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 53 kDa<br />Observed band size: 58 kDa<br /><br />Exposure time: 1 minute;<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/HA722069" style="font-weight: bold;text-decoration: underline;">HA722069</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Cdc25C on different lysates with Rabbit anti-Cdc25C antibody (HA722069) at 1/1,000 dilution.

    Lane 1: 293T cell lysate
    Lane 2: Caco-2 cell lysate
    Lane 3: HUVEC cell lysate
    Lane 4: HT-29 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 53 kDa
    Observed band size: 58 kDa

    Exposure time: 1 minute;

    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 (HA722069) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Cdc25C on different lysates with Rabbit anti-Cdc25C antibody (<a href="/products/HA722069" style="font-weight: bold;text-decoration: underline;">HA722069</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Cdc25C KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 58 kDa<br />Observed band size: 58 kDa<br /><br />Exposure time: 120 seconds; ECL: K1801;<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/HA722069" style="font-weight: bold;text-decoration: underline;">HA722069</a>) at 1/2,000 dilution was used in <a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a> at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of Cdc25C on different lysates with Rabbit anti-Cdc25C antibody (HA722069) at 1/2,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Cdc25C KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 58 kDa
    Observed band size: 58 kDa

    Exposure time: 120 seconds; ECL: K1801;

    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 (HA722069) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling Cdc25C with Rabbit anti-Cdc25C antibody (<a href="/products/HA722069" style="font-weight: bold;text-decoration: underline;">HA722069</a>) at 1/100 dilution.<br /><br />Cells were fixed in 100% precooled methanol 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-Cdc25C antibody (<a href="/products/HA722069" style="font-weight: bold;text-decoration: underline;">HA722069</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.

    Immunocytochemistry analysis of HeLa cells labeling Cdc25C with Rabbit anti-Cdc25C antibody (HA722069) at 1/100 dilution.

    Cells were fixed in 100% precooled methanol 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-Cdc25C antibody (HA722069) 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.

  • Flow cytometric analysis of HeLa cells labeling Cdc25C.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722069" style="font-weight: bold;text-decoration: underline;">HA722069</a>, 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&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 Cdc25C.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722069, 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 (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).

  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Cdc25C antibody (<a href="/products/HA722069" style="font-weight: bold;text-decoration: underline;">HA722069</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA722069" style="font-weight: bold;text-decoration: underline;">HA722069</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 human breast cancer tissue with Rabbit anti-Cdc25C antibody (HA722069) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA722069) 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