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  1. Link

    smyd3-antibody-epr19311-ab199361.pdf

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Epigenetics and Nuclear Signaling Chromatin Modifying Enzymes Methylation
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-SMYD3 antibody [EPR19311] (ab199361)

  • Datasheet
  • Certificate of Compliance
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Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)
  • Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)
  • Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)
  • Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)
  • Immunoprecipitation - Anti-SMYD3 antibody [EPR19311] (ab199361)
  • Immunoprecipitation - Anti-SMYD3 antibody [EPR19311] (ab199361)
  • Anti-SMYD3 antibody [EPR19311] (ab199361)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19311] to SMYD3
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-SMYD3 antibody [EPR19311]
    See all SMYD3 primary antibodies
  • Description

    Rabbit monoclonal [EPR19311] to SMYD3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human fetal kidney and fetal brain lysates; wild-type HAP1, HeLa, MCF7, HEK-293, T-47D, HCT 116, LLC, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain, kidney and spleen lysates; Rat brain and kidney lysates. IP: 293T whole cell lysate.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19311
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Methylation
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Polymerase associated factors
    • Pol II Transcription
    • Other
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Methylation
    • Lysine methylation
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Transcription factors

Associated products

  • Alternative Versions

    • Anti-SMYD3 antibody [EPR19311] - BSA and Azide free (ab251270)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab199361 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB
1/2000. Detects a band of approximately 45 kDa (predicted molecular weight: 49 kDa).
IP
1/50.
Notes
WB
1/2000. Detects a band of approximately 45 kDa (predicted molecular weight: 49 kDa).
IP
1/50.

Target

  • Function

    Histone methyltransferase. Specifically methylates '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' sequences.
  • Tissue specificity

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

    Belongs to the histone-lysine methyltransferase family.
    Contains 1 MYND-type zinc finger.
    Contains 1 SET domain.
  • Cellular localization

    Cytoplasm. Nucleus. Mainly cytoplasmic when cells are arrested at G0/G1. Accumulates in the nucleus at S phase and G2/M.
  • Target information above from: UniProt accession Q9H7B4 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 64754 Human
    • Entrez Gene: 69726 Mouse
    • Entrez Gene: 498295 Rat
    • Omim: 608783 Human
    • SwissProt: Q9H7B4 Human
    • SwissProt: Q9CWR2 Mouse
    • Unigene: 567571 Human
    • Unigene: 222338 Mouse
    • Alternative names

      • 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
      • Smyd3 antibody
      • SMYD3 protein antibody
      • SMYD3_HUMAN antibody
      • Zinc finger MYND domain containing 1 antibody
      • Zinc finger MYND domain containing protein 1 antibody
      • Zinc finger MYND domain-containing protein 1 antibody
      • Zinc finger protein subfamily 3A MYND domain containing 1 antibody
      • Zinc finger protein, subfamily 3A (MYND domain containing), 1 antibody
      • ZMYND 1 antibody
      • ZMYND1 antibody
      • ZNFN3A1 antibody
      see all

    Images

    • Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)
      Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)
      All lanes : Anti-SMYD3 antibody [EPR19311] (ab199361) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : SMYD3 knockout HAP1 whole cell lysate
      Lane 3 : HeLa whole cell lysate
      Lane 4 : MCF7 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

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



      Lanes 1 - 4: Merged signal (red and green). Green - ab199361 observed at 49 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

      ab199361 was shown to react with SMYD3 in HAP1 wild-type cells in Western blot. Loss of signal was observed when SMYD3 knockout sample was used. HAP1 wild-type and SMYD3 knockout whole cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab199361 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)
      Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)
      All lanes : Anti-SMYD3 antibody [EPR19311] (ab199361) at 1/2000 dilution

      Lane 1 : Human fetal kidney lysate
      Lane 2 : Human fetal brain lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

      Predicted band size: 49 kDa
      Observed band size: 45 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)
      Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)
      All lanes : Anti-SMYD3 antibody [EPR19311] (ab199361) at 1/2000 dilution

      Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
      Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 3 : T-47D (Human ductal breast epithelial tumor cell line) whole cell lysate
      Lane 4 : HCT 116 (Human colorectal carcinoma cell line) whole cell lysate
      Lane 5 : LLC (Mouse lung carcinoma cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 49 kDa
      Observed band size: 45 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)
      Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)
      All lanes : Anti-SMYD3 antibody [EPR19311] (ab199361) at 1/2000 dilution

      Lane 1 : Mouse brain lysate
      Lane 2 : Mouse kidney lysate
      Lane 3 : Mouse spleen lysate
      Lane 4 : Rat brain lysate
      Lane 5 : Rat kidney lysate
      Lane 6 : C6 (Rat glial tumor cell line) whole cell lysate
      Lane 7 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
      Lane 8 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
      Lane 9 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 49 kDa
      Observed band size: 45 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunoprecipitation - Anti-SMYD3 antibody [EPR19311] (ab199361)
      Immunoprecipitation - Anti-SMYD3 antibody [EPR19311] (ab199361)

      SMYD3 was immunoprecipitated from 1mg of 293T (Human epithelial cell line from embryonic kidney) whole cell lysate with ab199361 at 1/50 dilution.

      Western blot was performed from the immunoprecipitate using ab199361 at 1/1000 dilution.

      VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

      Lane 1: 293T whole cell lysate, 10µg (Input).

      Lane 2: ab199361 IP in 293T whole cell lysate.

      Lane 3: Rabbit IgG,monoclonal [EPR25A ]-Isotype Control (ab172730) instead of ab199361 in 293T whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • Immunoprecipitation - Anti-SMYD3 antibody [EPR19311] (ab199361)
      Immunoprecipitation - Anti-SMYD3 antibody [EPR19311] (ab199361)

      SMYD3 was immunoprecipitated from 1mg of 293T (Human epithelial cell line from embryonic kidney) whole cell lysate with ab199361 at 1/50 dilution.

      Western blot was performed from the immunoprecipitate using ab199361 at 1/1000 dilution (Panel A) or ab183498 at 1/1000 dilution (Panel B).

      VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

      Lane 1: 293T whole cell lysate, 10µg (Input).

      Lane 2: ab199361 IP in 293T whole cell lysate.

      Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab199361 in 293T whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 3 minutes.

      The product was validated with ab183498, which is specific to SMYD3.

    • Anti-SMYD3 antibody [EPR19311] (ab199361)
      Anti-SMYD3 antibody [EPR19311] (ab199361)

    Protocols

    • Western blot protocols
    • Immunoprecipitation protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (0)

    Publishing research using ab199361? Please let us know so that we can cite the reference in this datasheet.

    ab199361 has not yet been referenced specifically in any publications.

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