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Phospho-SMC1 (S957) Recombinant Rabbit Monoclonal Antibody [SC65-03]

Usd: 385

Specification

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Catalog# ET1610-55

Phospho-SMC1 (S957) Recombinant Rabbit Monoclonal Antibody [SC65-03]

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

Phospho-SMC1 (S957) Recombinant Rabbit Monoclonal Antibody [SC65-03]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser957 of Human SMC1 aa 945-980 / 1233.

Species Reactivity

Human

Validated Applications

WB, IHC-P, IF-Cell

Target Molecular Weight

Predicted band size: 144 kDa

Positive Control

HeLa cell lysate, HeLa treated with UV then recovery 30 minutes cell lysate, HeLa cells treated with UV then recovery 30 minutes, human tonsil tissue, human lung adenocarcinoma tissue, human breast carcinoma tissue.

Conjugation

unconjugated

Clone Number

SC65-03

RRID

AB_3073459

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:2,000

  • IHC-P

  • 1:50-1:200

  • IF-Cell

  • 1:50-1:200

Target

Function

The SMC (structural maintenance of chromosomes) family of proteins form heterodimeric complexes that modulate sister chromatid cohesion and chromosome condensation for mitosis. The two distinct classes of SMC protein complexes are comprised of SMC1 (also designated SB1.8) with SMC3 (also designated HCAP for human chromosome-associated protein and Bamacan for the secreted proteoglycan), and SMC2 (also designated hCAP-E) with SMC4 (also designated hCAP-C). The SMC1/SMC3 complex is required for metaphase progression in mitotic cells and functions independently of the SMC2/SMC4 complex during the cell cycle. SMC1 is ubiqitiously expressed in various human tissues, including thymus, testis and colon. SMC3 is expressed as a nuclear protein in the colon, but can also occur as a secreted proteoglycan expressed in testis and brain. The secreted proteoglycan contains several glycosylation sites and is thought to play a role in basement membrane physiology.

Background References

1. Qiu Z et al. Functional interactions between NURF and Ctcf regulate gene expression. Mol Cell Biol 35:224-37 (2015).

2. Mehta K et al. Human papillomaviruses activate and recruit SMC1 cohesin proteins for the differentiation-dependent life cycle through association with CTCF insulators. PLoS Pathog 11:e1004763 (2015).

Sequence Similarity

Belongs to the SMC family. SMC1 subfamily.

Post-translational Modification

Ubiquitinated by the DCX(DCAF15) complex, leading to its degradation.; Phosphorylated by ATM upon ionizing radiation in a NBS1-dependent manner. Phosphorylated by ATR upon DNA methylation in a MSH2/MSH6-dependent manner. Phosphorylation of Ser-957 and Ser-966 activates it and is required for S-phase checkpoint activation.

Subcellular Location

Nucleus, Chromosome, centromere.

UNIPROT

SwissProt: Q8NDV3 Human

SwissProt: Q14683 Human

Synonyms

CDLS2 antibody

DKFZp686L19178 antibody

DXhXs423e antibody

DXS423E antibody

KIAA0178 antibody

MGC138332 antibody

OTTHUMP00000061876 antibody

RP6 29D12.1 antibody

SB1.8 antibody

Segregation of mitotic chromosomes 1 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-SMC1 (S957) on different lysates with Rabbit anti-Phospho-SMC1 (S957) antibody (<a href="/products/ET1610-55" style="font-weight: bold;text-decoration: underline;">ET1610-55</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with UV then recovery 30 minutes cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 144 kDa<br />Observed band size: 150 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/ET1610-55" style="font-weight: bold;text-decoration: underline;">ET1610-55</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.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-SMC1 (S957) on different lysates with Rabbit anti-Phospho-SMC1 (S957) antibody (ET1610-55) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with UV then recovery 30 minutes cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 144 kDa
    Observed band size: 150 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 (ET1610-55) 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;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of HeLa cells treated with or without UV then recovery 30 minutes labeling Phospho-SMC1 (S957) with Rabbit anti-Phospho-SMC1 (S957) antibody (<a href="/products/ET1610-55" style="font-weight: bold;text-decoration: underline;">ET1610-55</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-SMC1 (S957) antibody (<a href="/products/ET1610-55" style="font-weight: bold;text-decoration: underline;">ET1610-55</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.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of HeLa cells treated with or without UV then recovery 30 minutes labeling Phospho-SMC1 (S957) with Rabbit anti-Phospho-SMC1 (S957) antibody (ET1610-55) 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-SMC1 (S957) antibody (ET1610-55) 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 tonsil tissue using anti-Phospho-SMC1 (S957) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1610-55" style="font-weight: bold;text-decoration: underline;">ET1610-55</a>, 1/50) for 30 minutes 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 using anti-Phospho-SMC1 (S957) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-55, 1/50) for 30 minutes 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 lung adenocarcinoma tissue using anti-Phospho-SMC1 (S957) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1610-55" style="font-weight: bold;text-decoration: underline;">ET1610-55</a>, 1/200) for 30 minutes 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 lung adenocarcinoma tissue using anti-Phospho-SMC1 (S957) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-55, 1/200) for 30 minutes 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 carcinoma tissue using anti-Phospho-SMC1 (S957) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1610-55" style="font-weight: bold;text-decoration: underline;">ET1610-55</a>, 1/50) for 30 minutes 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 carcinoma tissue using anti-Phospho-SMC1 (S957) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-55, 1/50) for 30 minutes 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.

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