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SMYD3 Recombinant Rabbit Monoclonal Antibody [JM73-63]

Usd: 385

Specification

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Catalog# ET1705-58

SMYD3 Recombinant Rabbit Monoclonal Antibody [JM73-63]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

SMYD3 Recombinant Rabbit Monoclonal Antibody [JM73-63]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human SMYD3 aa 251-300 / 428.

Species Reactivity

Human, Mouse, Rat

Validated Applications

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

Molecular Weight

Predicted band size: 49 kDa

Positive Control

MCF7 cell lysate, HCT 116 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, Mouse spleen tissue lysate, Rat spleen tissue lysate, Hela, HepG2, LOVO, human tonsil tissue, human colon cancer tissue, human placenta tissue, mouse stomach tissue.

Conjugation

unconjugated

Clone Number

JM73-63

RRID

AB_3070586

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

  • 1:500-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

  • IP

  • Use at an assay dependent concentration.

Target

Function

Belongs to the histone-lysine methyltransferase family. Contains 1 MYND-type zinc finger. Contains 1 SET domain. Histanes methyltransferase. Methylans 'Lys-4' of histone H3, inducing di- and tri-methylation, but not monomethylation. Plays an important role in transcriptional activation as a member of an RNA polymerase complex. Binds DNA containing 5'-CCCTCC- 3 'or 5'-GAGGGG-3'.

Background References

1. Liu T et al. Master redox regulator Trx1 upregulates SMYD1 & modulates lysine methylation. Biochim Biophys Acta 1854:1816-22 (2015).

2. Wang HF et al. Associations of the variable number of tandem repeats polymorphism in the SMYD3 gene with risk and prognosis of esophageal cancer: a case-control study. Neoplasma. doi: 10.4149/neo_2017_613 (2017).

Sequence Similarity

Belongs to the class V-like SAM-binding methyltransferase superfamily. Histone-lysine methyltransferase family.

Tissue Specificity

Expressed in skeletal muscles and testis. Overexpressed in a majority of colorectal and hepatocellular carcinomas.

Subcellular Location

Cytoplasm. Nucleus.

UNIPROT

SwissProt: Q9H7B4 Human

SwissProt: Q9CWR2 Mouse

Synonyms

bA74P14.1 (novel protein) antibody

bA74P14.1 antibody

FLJ21080 antibody

histone lysine N methyltransferase SMYD3 antibody

KMT3E antibody

MGC104324 antibody

SET and MYND domain containing 3 antibody

SET and MYND domain containing protein 3 antibody

SET and MYND domain-containing protein 3 antibody

SMYD 3 antibody

Expand

Images

  • Western blot analysis of SMYD3 on different lysates with Rabbit anti-SMYD3 antibody (<a href="/products/ET1705-58" style="font-weight: bold;text-decoration: underline;">ET1705-58</a>) at 1/2,000 dilution.<br /><br />Lane 1: MCF7 cell lysate (10 µg/Lane)<br />Lane 2: HCT 116 cell lysate (10 µg/Lane)<br />Lane 3: NIH/3T3 cell lysate (10 µg/Lane)<br />Lane 4: PC-12 cell lysate (10 µg/Lane)<br />Lane 5: Mouse spleen tissue lysate (20 µg/Lane)<br />Lane 6: Rat spleen tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 49 kDa<br />Observed band size: 40 kDa<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/ET1705-58" style="font-weight: bold;text-decoration: underline;">ET1705-58</a>) at 1/2,000 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 SMYD3 on different lysates with Rabbit anti-SMYD3 antibody (ET1705-58) at 1/2,000 dilution.

    Lane 1: MCF7 cell lysate (10 µg/Lane)
    Lane 2: HCT 116 cell lysate (10 µg/Lane)
    Lane 3: NIH/3T3 cell lysate (10 µg/Lane)
    Lane 4: PC-12 cell lysate (10 µg/Lane)
    Lane 5: Mouse spleen tissue lysate (20 µg/Lane)
    Lane 6: Rat spleen tissue lysate (20 µg/Lane)

    Predicted band size: 49 kDa
    Observed band size: 40 kDa

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-58) at 1/2,000 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 SMYD3 with Rabbit anti-SMYD3 antibody (<a href="/products/ET1705-58" style="font-weight: bold;text-decoration: underline;">ET1705-58</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-SMYD3 antibody (<a href="/products/ET1705-58" style="font-weight: bold;text-decoration: underline;">ET1705-58</a>) at 1/50 dilution in 2% negative goat serum 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 SMYD3 with Rabbit anti-SMYD3 antibody (ET1705-58) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-SMYD3 antibody (ET1705-58) at 1/50 dilution in 2% negative goat serum 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.

  • Immunocytochemistry analysis of HepG2 cells labeling SMYD3 with Rabbit anti-SMYD3 antibody (<a href="/products/ET1705-58" style="font-weight: bold;text-decoration: underline;">ET1705-58</a>) at 1/200 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-SMYD3 antibody (<a href="/products/ET1705-58" style="font-weight: bold;text-decoration: underline;">ET1705-58</a>) at 1/200 dilution in 2% negative goat serum 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 HepG2 cells labeling SMYD3 with Rabbit anti-SMYD3 antibody (ET1705-58) at 1/200 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-SMYD3 antibody (ET1705-58) at 1/200 dilution in 2% negative goat serum 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.

  • Immunocytochemistry analysis of LOVO cells labeling SMYD3 with Rabbit anti-SMYD3 antibody (<a href="/products/ET1705-58" style="font-weight: bold;text-decoration: underline;">ET1705-58</a>) at 1/200 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-SMYD3 antibody (<a href="/products/ET1705-58" style="font-weight: bold;text-decoration: underline;">ET1705-58</a>) at 1/200 dilution in 2% negative goat serum 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 SMYD3 with Rabbit anti-SMYD3 antibody (ET1705-58) at 1/200 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-SMYD3 antibody (ET1705-58) at 1/200 dilution in 2% negative goat serum 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..

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-SMYD3 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-SMYD3 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-SMYD3 antibody (<a href="/products/ET1705-58" style="font-weight: bold;text-decoration: underline;">ET1705-58</a>) at 1/50 dilution.Hela cells<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/ET1705-58" style="font-weight: bold;text-decoration: underline;">ET1705-58</a>) at 1/50 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 colon cancer tissue with Rabbit anti-SMYD3 antibody (ET1705-58) at 1/50 dilution.Hela cells
    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 (ET1705-58) at 1/50 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 placenta tissue with Rabbit anti-SMYD3 antibody (<a href="/products/ET1705-58" style="font-weight: bold;text-decoration: underline;">ET1705-58</a>) at 1/50 dilution.Hela cells<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/ET1705-58" style="font-weight: bold;text-decoration: underline;">ET1705-58</a>) at 1/50 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 placenta tissue with Rabbit anti-SMYD3 antibody (ET1705-58) at 1/50 dilution.Hela cells
    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 (ET1705-58) at 1/50 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 stomach tissue with Rabbit anti-SMYD3 antibody (<a href="/products/ET1705-58" style="font-weight: bold;text-decoration: underline;">ET1705-58</a>) at 1/50 dilution.Hela cells<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/ET1705-58" style="font-weight: bold;text-decoration: underline;">ET1705-58</a>) at 1/50 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 stomach tissue with Rabbit anti-SMYD3 antibody (ET1705-58) at 1/50 dilution.Hela cells
    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 (ET1705-58) at 1/50 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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