Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)

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Overview

  • Product name

    Anti-KMT1B / SUV39H2 antibody [EPR18495]
    See all KMT1B / SUV39H2 primary antibodies

  • Description

    Rabbit monoclonal [EPR18495] to KMT1B / SUV39H2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, IP, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human KMT1B/ SUV39H2 aa 350 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9H5I1

  • Positive control

    • WB: HEK-293, HeLa, NIH/3T3, SH-SY5Y and MOLT-4 cell lysates; human fetal kidney lysate; human, mouse and rat testis lysates. IHC-P: Human, mouse and rat testis tissues. IP: SH-SY5Y 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 rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab190870 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
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/50.
WB 1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).

Target

  • Function

    Histone methyltransferase that specifically trimethylates 'Lys-9' of histone H3 using monomethylated H3 'Lys-9' as substrate. H3 'Lys-9' trimethylation represents a specific tag for epigenetic transcriptional repression by recruiting HP1 (CBX1, CBX3 and/or CBX5) proteins to methylated histones. Mainly functions in heterochromatin regions, thereby playing a central role in the establishment of constitutive heterochromatin at pericentric and telomere regions. H3 'Lys-9' trimethylation is also required to direct DNA methylation at pericentric repeats. SUV39H1 is targeted to histone H3 via its interaction with RB1 and is involved in many processes, such as cell cycle regulation, transcriptional repression and regulation of telomere length. May participate in regulation of higher order chromatin organization during spermatogenesis.

  • Sequence similarities

    Belongs to the histone-lysine methyltransferase family. Suvar3-9 subfamily.
    Contains 1 chromo domain.
    Contains 1 post-SET domain.
    Contains 1 pre-SET domain.
    Contains 1 SET domain.

  • Domain

    Although the SET domain contains the active site of enzymatic activity, both pre-SET and post-SET domains are required for methyltransferase activity. The SET domain also participates to stable binding to heterochromatin.

  • Cellular localization

    Nucleus. Chromosome > centromere. Associates with centromeric constitutive heterochromatin.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • FLJ23414 antibody
    • H3 K9 HMTase 2 antibody
    • H3-K9-HMTase 2 antibody
    • Histone H3 K9 methyltransferase 2 antibody
    • Histone H3-K9 methyltransferase 2 antibody
    • Histone lysine N methyltransferase H3 lysine 9 specific 2 antibody
    • Histone lysine N methyltransferase SUV39H2 antibody
    • Histone-lysine N-methyltransferase SUV39H2 antibody
    • KMT1B antibody
    • Lysine N methyltransferase 1B antibody
    • Lysine N-methyltransferase 1B antibody
    • sSuppressor of variegation 3 9 homolog 2 (Drosophila) antibody
    • Su(var)3 9 Drosophila homolog of 2 antibody
    • Su(var)3 9 homolog 2 antibody
    • Su(var)3-9 homolog 2 antibody
    • Suppressor of variegation 3 9 homolog 2 antibody
    • Suppressor of variegation 3-9 homolog 2 antibody
    • Suv39h2 antibody
    • SUV92_HUMAN antibody
    see all

Images

  • Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)
    Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)
    All lanes : Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870) at 1/1000 dilution

    Lane 1 : Wild-type U2-OS whole cell lysate
    Lane 2 : SUV39H2 knockout U2-OS whole cell lysate
    Lane 3 : HEK-293 whole cell lysate
    Lane 4 : HeLa whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 47 kDa
    Observed band size: 47 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab190870 observed at 47 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab190870 was shown to recognize SUV39H2 in wild-type U2-OS cells as signal was lost at the expected MW in SUV39H2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SUV39H2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab190870 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)
    Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)
    All lanes : Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870) at 1/1000 dilution

    Lane 1 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate. at 20 µg
    Lane 2 : MOLT-4 (Human lymphoblastic leukemia cell line) whole cell lysate at 20 µg
    Lane 3 : Human fetal kidney at 10 µg

    Secondary
    Lanes 1-2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
    Lane 3 : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Developed using the ECL technique.

    Predicted band size: 47 kDa
    Observed band size: 47 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST

  • Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)
    Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)
    All lanes : Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870) at 1/10000 dilution

    Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cell line) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Developed using the ECL technique.

    Predicted band size: 47 kDa
    Observed band size: 47,54 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The full length SUV39H2 orthologues differ in size: human 410aa (UniProt Q9H5I1), mouse 477aa (UniProt Q9EQQ0) and rat 481aa (UniProt D3ZIH5).

  • Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)
    Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)
    All lanes : Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870) at 1/5000 dilution

    Lane 1 : Human testis lysate
    Lane 2 : Mouse testis lysate
    Lane 3 : Rat testis lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Developed using the ECL technique.

    Predicted band size: 47 kDa
    Observed band size: 47,54 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The full length SUV39H2 orthologues differ in size: human 410aa (UniProt Q9H5I1), mouse 477aa (UniProt Q9EQQ0) and rat 481aa (UniProt D3ZIH5).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)

    Immunohistochemical analysis of paraffin-embedded human testis tissue labeling KMT1B / SUV39H2 with ab190870 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear staining on germ cells of human testis is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling KMT1B / SUV39H2 with ab190870 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nucleuarstaining on germ cells of mouse testis is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)

    Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling KMT1B / SUV39H2 with ab190870 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear staining on germ cells of rat testis is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)
    Immunoprecipitation - Anti-KMT1B / SUV39H2 antibody [EPR18495] (ab190870)

    KMT1B / SUV39H2 was immunoprecipitated from 1mg of SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate with ab190870 at 1/50 dilution.

    Western blot was performed from the immunoprecipitate using ab190870 at 1/1000 dilution.

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

    Lane 1: SH-SY5Y whole cell lysate 10µg (Input).

    Lane 2: ab190870 IP in SH-SY5Y whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab190870 in SH-SY5Y whole cell lysate.

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

    Exposure time: 30 seconds.

References

This product has been referenced in:

  • Shuai W  et al. SUV39H2 promotes colorectal cancer proliferation and metastasis via tri-methylation of the SLIT1 promoter. Cancer Lett 422:56-69 (2018). WB, IHC . Read more (PubMed: 29458143) »
See 1 Publication for this product

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