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Phospho-MCM2 (S27) Recombinant Rabbit Monoclonal Antibody [JE63-40]

Usd: 385

Specification

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Catalog# HA722574

Phospho-MCM2 (S27) Recombinant Rabbit Monoclonal Antibody [JE63-40]

Application

  • WB

  • IF-Cell

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

Phospho-MCM2 (S27) Recombinant Rabbit Monoclonal Antibody [JE63-40]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser27 of Human MCM2.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, IP

Molecular Weight

Predicted band size: 102 kDa

Positive Control

HeLa cell lysate, Jurkat cell lysate, NIH/3T3 cell lysate, Mouse thymus tissue lysate, Rat thymus tissue lysate, HeLa, human cervical carcinoma tissue, human lymph node tissue, mouse lymph node tissue, rat lymph node tissue.

Conjugation

unconjugated

Clone Number

JE63-40

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

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:1,000-1:3,000

  • IP

  • 1-2μg/sample

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. Sun Y et al. MCM2 in human cancer: functions, mechanisms, and clinical significance. Mol Med. 2022 Oct

2. Zhou X et al. MCM2 promotes the stemness and sorafenib resistance of hepatocellular carcinoma cells via hippo signaling. Cell Death Discov. 2022 Oct

Subcellular Location

Nucleus.

UNIPROT

SwissProt: P49736 Human

SwissProt: P97310 Mouse

Entrez Gene: 312538 Rat

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

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-MCM2 (S27) on different lysates with Rabbit anti-Phospho-MCM2 (S27) antibody (<a href="/products/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</a>) at 1/1,000 dilution and pan MCM2 antibody (<a href="/products/HA721553" style="font-weight: bold;text-decoration: underline;">HA721553</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: Jurkat cell lysate (20 µg/Lane)<br />Lane 3: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 4: Mouse thymus tissue lysate (40 µg/Lane)<br />Lane 5: Rat thymus tissue lysate (40 µg/Lane)<br />Lane 6: HeLa cell lysate, the membrane treated with λpp for 1 hour (20 µg/Lane)<br /><br />Predicted band size: 102 kDa<br />Observed band size: 140 kDa<br /><br />Exposure time: 17 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/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</a>) at 1/1,000 dilution and pan MCM2 antibody (<a href="/products/HA721553" style="font-weight: bold;text-decoration: underline;">HA721553</a>) at 1/1,000 dilution were 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.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-MCM2 (S27) on different lysates with Rabbit anti-Phospho-MCM2 (S27) antibody (HA722574) at 1/1,000 dilution and pan MCM2 antibody (HA721553) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: Jurkat cell lysate (20 µg/Lane)
    Lane 3: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 4: Mouse thymus tissue lysate (40 µg/Lane)
    Lane 5: Rat thymus tissue lysate (40 µg/Lane)
    Lane 6: HeLa cell lysate, the membrane treated with λpp for 1 hour (20 µg/Lane)

    Predicted band size: 102 kDa
    Observed band size: 140 kDa

    Exposure time: 17 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 (HA722574) at 1/1,000 dilution and pan MCM2 antibody (HA721553) at 1/1,000 dilution were 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.

  • Immunocytochemistry analysis of HeLa cells labeling Phospho-MCM2 (S27) with Rabbit anti-Phospho-MCM2 (S27) antibody (<a href="/products/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</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-Phospho-MCM2 (S27) antibody (<a href="/products/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</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 Phospho-MCM2 (S27) with Rabbit anti-Phospho-MCM2 (S27) antibody (HA722574) 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-Phospho-MCM2 (S27) antibody (HA722574) 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.

  • Immunohistochemical analysis of paraffin-embedded human cervical carcinoma tissue with Rabbit anti-Phospho-MCM2 (S27) antibody (<a href="/products/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</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/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</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 cervical carcinoma tissue with Rabbit anti-Phospho-MCM2 (S27) antibody (HA722574) 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 (HA722574) 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 lymph node tissue with Rabbit anti-Phospho-MCM2 (S27) antibody (<a href="/products/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</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/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</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 lymph node tissue with Rabbit anti-Phospho-MCM2 (S27) antibody (HA722574) 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 (HA722574) 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 mouse lymph node tissue with Rabbit anti-Phospho-MCM2 (S27) antibody (<a href="/products/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</a>) at 1/3,000 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/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</a>) at 1/3,000 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 lymph node tissue with Rabbit anti-Phospho-MCM2 (S27) antibody (HA722574) at 1/3,000 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 (HA722574) at 1/3,000 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 lymph node tissue with Rabbit anti-Phospho-MCM2 (S27) antibody (<a href="/products/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</a>) at 1/3,000 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/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</a>) at 1/3,000 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 lymph node tissue with Rabbit anti-Phospho-MCM2 (S27) antibody (HA722574) at 1/3,000 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 (HA722574) at 1/3,000 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.

  • Phospho-MCM2 (S27) was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HeLa cell lysate (input)<br />Lane 2: <a href="/products/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA722574" style="font-weight: bold;text-decoration: underline;">HA722574</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 20 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    Phospho-MCM2 (S27) was immunoprecipitated from 0.2 mg HeLa cell lysate with HA722574 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722574 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: HA722574 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of HA722574 in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 20 seconds; ECL: K1801

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