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

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

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18495] to KMT1B / SUV39H2
Suitable for: IHC-P, IP, WB
Knockout validated
Reacts with: Mouse, Rat, Human

Carrier Free

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

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, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18495
Isotype

IgG
Research areas

Alternative Versions
- Anti-KMT1B / SUV39H2 antibody [EPR18495] - BSA and Azide free (ab240313)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human SUV39H2 knockout U-2 OS cell line (ab261882)
KO cell lysates
- Human SUV39H2 knockout U-2 OS cell lysate (ab261691)
Recombinant Protein
- Recombinant human KMT1B / SUV39H2 protein (ab80288)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

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).

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).

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.
Target information above from: UniProt accession Q9H5I1 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 79723 Human
- Entrez Gene: 64707 Mouse
- Entrez Gene: 364785 Rat
- Omim: 606503 Human
- SwissProt: Q9H5I1 Human
- SwissProt: Q9EQQ0 Mouse
- Unigene: 554883 Human
- Unigene: 128273 Mouse
- Unigene: 23483 Mouse
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

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)

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)

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)

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)

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)

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)

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)

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

Western blot protocols
Immunoprecipitation protocols
Immunohistochemistry protocols

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab190870? Please let us know so that we can cite the reference in this datasheet.

ab190870 has been referenced in 4 publications.

Miao Y et al. Histone methyltransferase SUV39H2 regulates LSD1-dependent CDH1 expression and promotes epithelial mesenchymal transition of osteosarcoma. Cancer Cell Int 21:2 (2021). PubMed: 33397384
Jing J et al. The histone methyltransferase Suv39h regulates 3T3-L1 adipogenesis. Adipocyte 9:401-414 (2020). PubMed: 32698678
Grigore F et al. BRAF inhibition in melanoma is associated with the dysregulation of histone methylation and histone methyltransferases. Neoplasia 22:376-389 (2020). PubMed: 32629178
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 . PubMed: 29458143

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Domain

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (4)

Customer reviews and Q&As