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Sumo 2+3 Recombinant Rabbit Monoclonal Antibody [JJ087-04]

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Specification

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Catalog# ET1701-17

Sumo 2+3 Recombinant Rabbit Monoclonal Antibody [JJ087-04]

Application

  • WB

  • IF-Cell

  • 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

Sumo 2+3 Recombinant Rabbit Monoclonal Antibody [JJ087-04]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Sumo 3 aa 11-60 / 103.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P

Molecular Weight

Predicted band size: 12 kDa

Positive Control

HeLa cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, Human lung tissue lysate, Mouse kidney tissue lysate, HeLa, NIH/3T3, human kidney tissue, mouse kidney tissue, rat kidney tissue, mouse large intestine tissue, rat skin tissue.

Conjugation

unconjugated

Clone Number

JJ087-04

RRID

AB_3070184

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. 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

  • 1:500-1:1,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:50-1:1,000

Target

Function

The small ubiquitin-related modifier (SUMO) proteins, which include SUMO-1, SUMO-2 and SUMO-3, belong to the ubiquitin-like protein family. Like ubiquitin, the SUMO proteins are synthesized as precursor proteins that undergo processing before conjugation to target proteins. Also, both utilize the E1, E2, and E3 cascade enzymes for conjugation. However, SUMO and ubiquitin differ with respect to targeting. Ubiquitination predominantly targets proteins for degradation, whereas sumoylation targets proteins to a variety of cellular processing, including nuclear transport, transcriptional regulation, apoptosis and protein stability. The unconjugated SUMO-1, SUMO-2 and SUMO-3 proteins localize to the nuclear membrane, nuclear bodies and cytoplasm, respectively. SUMO-1 utilizes Ubc9 for conjugation to several target proteins, which include IkBa, MDM2, p53, PML and Ran GAP1. SUMO-2 and SUMO-3 contribute to a greater percentage of protein modification than does SUMO-1, and unlike SUMO-1, they can form polymeric chains. In addition, SUMO-3 regulates b-Amyloid generation and may be critical in the onset or progression of Alzheimer's disease.

Background References

1. Sloan E et al. Analysis of the SUMO2 Proteome during HSV-1 Infection. PLoS Pathog 11:e1005059 (2015).

2. Conn KL et al. Novel Role for Protein Inhibitor of Activated STAT 4 (PIAS4) in the Restriction of Herpes Simplex Virus 1 by the Cellular Intrinsic Antiviral Immune Response. J Virol 90:4807-26 (2016).

Sequence Similarity

Belongs to the ubiquitin family. SUMO subfamily.

Tissue Specificity

Expressed predominantly in liver.

Post-translational Modification

Polymeric chains can be formed through Lys-11 cross-linking.; Cleavage of precursor form by SENP1, SENP2 or SENP5 is necessary for function.

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: P55854 Human

SwissProt: P61956 Human

SwissProt: P61957 Mouse

SwissProt: Q9Z172 Mouse

SwissProt: P61959 Rat

SwissProt: Q5XIF4 Rat

Synonyms

HSMT3 antibody

MGC117191 antibody

OTTHUMP00000115275 antibody

OTTHUMP00000115276 antibody

OTTHUMP00000115277 antibody

Sentrin 2 antibody

Small ubiquitin like modifier 2 antibody

Small ubiquitin related modifier 2 antibody

small ubiquitin-like modifier 3 antibody

small ubiquitin-related modifier 3 antibody

Expand

Images

  • Western blot analysis of Sumo 2+3 on different lysates with Rabbit anti-Sumo 2+3 antibody (<a href="/products/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 3: PC-12 cell lysate (20 µg/Lane)<br />Lane 4: Human lung tissue lysate (40 µg/Lane)<br />Lane 5: Mouse kidney tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 12 kDa<br />Observed band size: 16 kDa<br /><br />Exposure time: 2 minutes; ECL: K1802;<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/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</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 Sumo 2+3 on different lysates with Rabbit anti-Sumo 2+3 antibody (ET1701-17) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 3: PC-12 cell lysate (20 µg/Lane)
    Lane 4: Human lung tissue lysate (40 µg/Lane)
    Lane 5: Mouse kidney tissue lysate (40 µg/Lane)

    Predicted band size: 12 kDa
    Observed band size: 16 kDa

    Exposure time: 2 minutes; ECL: K1802;

    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 (ET1701-17) 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.

  • Immunocytochemistry analysis of HeLa cells labeling Sumo 2+3 with Rabbit anti-Sumo 2+3 antibody (<a href="/products/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</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-Sumo 2+3 antibody (<a href="/products/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</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/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</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 Sumo 2+3 with Rabbit anti-Sumo 2+3 antibody (ET1701-17) 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-Sumo 2+3 antibody (ET1701-17) 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 (HA601187, 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.

  • Immunocytochemistry analysis of NIH/3T3 cells labeling Sumo 2+3 with Rabbit anti-Sumo 2+3 antibody (<a href="/products/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</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-Sumo 2+3 antibody (<a href="/products/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</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/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</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 NIH/3T3 cells labeling Sumo 2+3 with Rabbit anti-Sumo 2+3 antibody (ET1701-17) 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-Sumo 2+3 antibody (ET1701-17) 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 (HA601187, 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 kidney tissue with Rabbit anti-Sumo 2+3 antibody (<a href="/products/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</a>) at 1/200 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/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</a>) at 1/200 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 kidney tissue with Rabbit anti-Sumo 2+3 antibody (ET1701-17) at 1/200 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 (ET1701-17) at 1/200 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 kidney tissue with Rabbit anti-Sumo 2+3 antibody (<a href="/products/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</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/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</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 mouse kidney tissue with Rabbit anti-Sumo 2+3 antibody (ET1701-17) 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 (ET1701-17) 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 rat kidney tissue with Rabbit anti-Sumo 2+3 antibody (<a href="/products/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</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/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</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 rat kidney tissue with Rabbit anti-Sumo 2+3 antibody (ET1701-17) 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 (ET1701-17) 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 large intestine tissue with Rabbit anti-Sumo 2+3 antibody (<a href="/products/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</a>) at 1/400 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/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</a>) at 1/400 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 large intestine tissue with Rabbit anti-Sumo 2+3 antibody (ET1701-17) at 1/400 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 (ET1701-17) at 1/400 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 skin tissue with Rabbit anti-Sumo 2+3 antibody (<a href="/products/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</a>) at 1/400 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/ET1701-17" style="font-weight: bold;text-decoration: underline;">ET1701-17</a>) at 1/400 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 skin tissue with Rabbit anti-Sumo 2+3 antibody (ET1701-17) at 1/400 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 (ET1701-17) at 1/400 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.

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