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MCM2 Recombinant Rabbit Monoclonal Antibody [PD00-90]

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Specification

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Catalog# HA721177

MCM2 Recombinant Rabbit Monoclonal Antibody [PD00-90]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

Conjugation

  • unconjugated

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

Overview

Product Name

MCM2 Recombinant Rabbit Monoclonal Antibody [PD00-90]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within

Species Reactivity

Human

Validated Applications

WB, IHC-P, IF-Cell, FC

Molecular Weight

Predicted band size: 102 kDa

Positive Control

HL-60 cell lysate, SiHa cell lysate, human tonsil tissue, human cervical carcinoma tissue, human ovarian carcinoma tissue, human small intestine tissue, LoVo, HL-60.

Conjugation

unconjugated

Clone Number

PD00-90

RRID

AB_3072297

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

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:800-1:2,000

  • IF-Cell

  • 1:100

  • FC

  • 1:500-1:1,000

Target

Function

DNA replication licensing factor MCM2 is a protein that in humans is encoded by the MCM2 gene. The protein encoded by this gene is one of the highly conserved mini-chromosome maintenance proteins (MCM) that are involved in the initiation of eukaryotic genome replication. The hexameric protein complex formed by MCM proteins is a key component of the pre-replication complex (pre-RC) and may be involved in the formation of replication forks and in the recruitment of other DNA replication related proteins. This protein forms a complex with MCM4, 6, and 7, and has been shown to regulate the helicase activity of the complex. This protein is phosphorylated, and thus regulated by, protein kinases CDC2 and CDC7.

Background References

1. Keyhanian K et al. Combination of MCM2 With Ki67 and p16 Immunohistochemistry Can Distinguish Uterine Leiomyosarcomas. Int J Gynecol Pathol. 2020 Jul

2. Del Moral-Hernández O et al. TOP2A/MCM2, p16INK4a, and cyclin E1 expression in liquid-based cytology: a biomarkers panel for progression risk of cervical premalignant lesions. BMC Cancer. 2021 Jan

Subcellular Location

Nucleus, Chromosome.

UNIPROT

SwissProt: P49736 Human

Synonyms

BM28 antibody

CCNL 1 antibody

CCNL1 antibody

CDC like 1 antibody

cdc19 antibody

CDCL 1 antibody

CDCL1 antibody

Cell devision cycle like 1 antibody

Cyclin like 1 antibody

D3S3194 antibody

Expand

Images

  • Western blot analysis of MCM2 on different lysates with Rabbit anti-MCM2 antibody (<a href="/products/HA721177" style="font-weight: bold;text-decoration: underline;">HA721177</a>) at 1/2,000 dilution.<br /><br />Lane 1: HL-60 cell lysate<br />Lane 2: SiHa cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 102 kDa<br />Observed band size: 130 kDa<br /><br />Exposure time: 1 minute;<br /><br />8% 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/HA721177" style="font-weight: bold;text-decoration: underline;">HA721177</a>) at 1/2,000 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 1:300,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of MCM2 on different lysates with Rabbit anti-MCM2 antibody (HA721177) at 1/2,000 dilution.

    Lane 1: HL-60 cell lysate
    Lane 2: SiHa cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 102 kDa
    Observed band size: 130 kDa

    Exposure time: 1 minute;

    8% 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 (HA721177) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-MCM2 antibody (<a href="/products/HA721177" style="font-weight: bold;text-decoration: underline;">HA721177</a>) at 1/1,000 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/HA721177" style="font-weight: bold;text-decoration: underline;">HA721177</a>) at 1/1,000 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 tonsil tissue with Rabbit anti-MCM2 antibody (HA721177) at 1/1,000 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 (HA721177) at 1/1,000 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 cervical carcinoma tissue with Rabbit anti-MCM2 antibody (<a href="/products/HA721177" style="font-weight: bold;text-decoration: underline;">HA721177</a>) at 1/800 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/HA721177" style="font-weight: bold;text-decoration: underline;">HA721177</a>) at 1/800 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 cervical carcinoma tissue with Rabbit anti-MCM2 antibody (HA721177) at 1/800 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 (HA721177) at 1/800 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 ovarian carcinoma tissue with Rabbit anti-MCM2 antibody (<a href="/products/HA721177" style="font-weight: bold;text-decoration: underline;">HA721177</a>) at 1/1,000 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/HA721177" style="font-weight: bold;text-decoration: underline;">HA721177</a>) at 1/1,000 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 ovarian carcinoma tissue with Rabbit anti-MCM2 antibody (HA721177) at 1/1,000 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 (HA721177) at 1/1,000 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 small intestine tissue with Rabbit anti-MCM2 antibody (<a href="/products/HA721177" style="font-weight: bold;text-decoration: underline;">HA721177</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/HA721177" style="font-weight: bold;text-decoration: underline;">HA721177</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 small intestine tissue with Rabbit anti-MCM2 antibody (HA721177) 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 (HA721177) 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.

  • Immunocytochemistry analysis of LoVo cells labeling MCM2 with Rabbit anti-MCM2 antibody (<a href="/products/HA721177" style="font-weight: bold;text-decoration: underline;">HA721177</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-MCM2 antibody (<a href="/products/HA721177" style="font-weight: bold;text-decoration: underline;">HA721177</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 LoVo cells labeling MCM2 with Rabbit anti-MCM2 antibody (HA721177) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-MCM2 antibody (HA721177) 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 HL-60 cells labeling MCM2.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721177" style="font-weight: bold;text-decoration: underline;">HA721177</a>, 1ug/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 HL-60 cells labeling MCM2.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721177, 1ug/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).

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Citation