Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-SMYD3 antibody [EPR19311] (ab199361)

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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 within Human SMYD3 aa 1-250. The exact sequence is proprietary.
    Database link: Q9H7B4

  • 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

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

    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.

Properties

Applications

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.

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.
  • Information by UniProt

  • Database links

    see all

  • 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.

References

ab199361 has not yet been referenced specifically in any publications.

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

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