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Rabbit Recombinant Monoclonal NSUN2/SAKI antibody. Suitable for ICC/IF, IP, WB, Flow Cyt (Intra), IHC-P and reacts with Mouse, Human, Rat samples. Cited in 2 publications.

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Images

Immunoprecipitation - Anti-NSUN2/SAKI antibody [EPR24140-93] (AB259941), expandable thumbnail
  • Western blot - Anti-NSUN2/SAKI antibody [EPR24140-93] (AB259941), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-NSUN2/SAKI antibody [EPR24140-93] (AB259941), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSUN2/SAKI antibody [EPR24140-93] (AB259941), expandable thumbnail
  • Immunoprecipitation - Anti-NSUN2/SAKI antibody [EPR24140-93] (AB259941), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ICC/IFIPWBFlow Cyt (Intra)IHC-P
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Tested
Tested
Tested
Tested
Tested
Rat
Expected
Expected
Tested
Expected
Tested

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

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

Species

Human

Dilution info

1/1000

Notes

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

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/30

Notes

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

Species

Human

Dilution info

1/30

Notes

-

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

-

Species

Rat

Dilution info

1/1000

Notes

-

Species

Human

Dilution info

1/1000

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/50

Notes

Use at 1/500 dilution for HeLa.

Species

Human

Dilution info

1/50

Notes

Use at 1/500 dilution for HeLa.

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/200

Notes

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

Species

Rat

Dilution info

1/200

Notes

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

Species

Human

Dilution info

1/200

Notes

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

Target data

Function

RNA cytosine C(5)-methyltransferase that methylates cytosine to 5-methylcytosine (m5C) in various RNAs, such as tRNAs, mRNAs and some long non-coding RNAs (lncRNAs) (PubMed:17071714, PubMed:22995836, PubMed:31199786, PubMed:31358969). Involved in various processes, such as epidermal stem cell differentiation, testis differentiation and maternal to zygotic transition during early development: acts by increasing protein synthesis; cytosine C(5)-methylation promoting tRNA stability and preventing mRNA decay (PubMed:31199786). Methylates cytosine to 5-methylcytosine (m5C) at positions 34 and 48 of intron-containing tRNA(Leu)(CAA) precursors, and at positions 48, 49 and 50 of tRNA(Gly)(GCC) precursors (PubMed:17071714, PubMed:22995836, PubMed:31199786). tRNA methylation is required generation of RNA fragments derived from tRNAs (tRFs) (PubMed:31199786). Also mediates C(5)-methylation of mitochondrial tRNAs (PubMed:31276587). Catalyzes cytosine C(5)-methylation of mRNAs, leading to stabilize them and prevent mRNA decay: mRNA stabilization involves YBX1 that specifically recognizes and binds m5C-modified transcripts (PubMed:22395603, PubMed:31358969, PubMed:34556860). Cytosine C(5)-methylation of mRNAs also regulates mRNA export: methylated transcripts are specifically recognized by THOC4/ALYREF, which mediates mRNA nucleo-cytoplasmic shuttling (PubMed:28418038). Also mediates cytosine C(5)-methylation of non-coding RNAs, such as vault RNAs (vtRNAs), promoting their processing into regulatory small RNAs (PubMed:23871666). Cytosine C(5)-methylation of vtRNA VTRNA1.1 promotes its processing into small-vault RNA4 (svRNA4) and regulates epidermal differentiation (PubMed:31186410). May act downstream of Myc to regulate epidermal cell growth and proliferation (By similarity). Required for proper spindle assembly and chromosome segregation, independently of its methyltransferase activity (PubMed:19596847).

Alternative names

Recommended products

Rabbit Recombinant Monoclonal NSUN2/SAKI antibody. Suitable for ICC/IF, IP, WB, Flow Cyt (Intra), IHC-P and reacts with Mouse, Human, Rat samples. Cited in 2 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR24140-93

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

NSUN2 also known as SAKI is a methyltransferase enzyme that adds a methyl group to cytosine within nucleic acids most often in RNA. It has a molecular mass of about 83 kDa. This protein is widely expressed in different tissues but shows high expression levels in the brain and reproductive organs. NSUN2 modifies RNA molecules impacting their function and stability which is essential in various cellular processes.

Biological function summary

NSUN2 participates in RNA metabolism and processing. By introducing m5C (5-methylcytosine) modifications to RNA NSUN2 influences RNA stability transport and translation. This protein is a part of larger complexes that regulate diverse cellular functions and maintains transcriptome integrity. Alterations in NSUN2 activity can affect cellular proliferation and differentiation highlighting its role in maintaining normal cell function.

Pathways

NSUN2 plays an integral role in the epitranscriptomic regulation pathway in particular RNA modification and gene expression regulation pathways. It acts alongside proteins like TRMT2A and TRMT61A which are involved in similar processes of RNA methylation and modification. These pathways help maintain a balance in gene expression profiles necessary for proper cellular function and response to external signals.

Associated diseases and disorders

Mutations or dysfunctions in NSUN2 are associated with mental retardation and cancer. In neurodevelopmental disorders altered NSUN2 activity disrupts normal brain development potentially linking it with proteins like FTSJ1 another RNA methyltransferase. Similarly in cancer NSUN2 dysregulation leads to aberrant cell proliferation and tumorigenesis often in connection with oncogenic pathways and other modified proteins.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

12 product images

  • Immunoprecipitation - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941), expandable thumbnail

    Immunoprecipitation - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941)

    NSUN2/SAKI was immunoprecipitated from 0.35 mg HEK-293 (human embryonic kidney epithelial cell) whole cell lysate with ab259941 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259941 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 10 ug

    Lane 2: ab259941 IP in HEK-293 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab259941 in HEK-293 whole cell lysate

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

    Exposure time: 15 seconds.

    All lanes: Immunoprecipitation - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941)

    Predicted band size: 86 kDa

    Observed band size: 100 kDa

  • Western blot - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941), expandable thumbnail

    Western blot - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941)

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

    The observed MW is consistent with what has described in the literature (PMID: 23604283, 25233213, 27447970, 31487418).

    Exposure times: Lane 1: 15 seconds

    Lane 2-4: 48 seconds.

    All lanes: Western blot - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941) at 1/1000 dilution

    Lane 1: HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 3: HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

    Lane 4: A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

    Secondary

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

    Predicted band size: 86 kDa

    Observed band size: 100 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labelling NSUN2/SAKI with ab259941 at 1/1000 dilution (0.503 μg/ml), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor®488) antibody at 1/1000 dilution (Green). Confocal image showing weak cytoplasmic and strong nuclear staining in HeLa cells. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941)

    Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling NSUN2/SAKI with ab259941 at 1/200 dilution (2.515 μg/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Nuclear staining on human pancreas (PMID: 16713953). The section was incubated with ab259941 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunoprecipitation - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941), expandable thumbnail

    Immunoprecipitation - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941)

    NSUN2/SAKI was immunoprecipitated from 0.35 mg mouse spleen tissue lysate with ab259941 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259941 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: Mouse spleen tissue lysate 10 ug

    Lane 2: ab259941 IP in Mouse spleen tissue lysate

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab259941 in mouse spleen tissue lysate

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

    Exposure time: 15 seconds.

    All lanes: Immunoprecipitation - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941)

    Predicted band size: 86 kDa

    Observed band size: 100 kDa

  • Western blot - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941), expandable thumbnail

    Western blot - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941)

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

    The observed MW is consistent with what has described in the literature (PMID: 27306184).

    Exposure times: Lane 1: 48 seconds

    Lane 2, 3: 3 minutes.

    All lanes: Western blot - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941) at 1/1000 dilution

    Lane 1: ES-D3 (mouse embryonic mtipotent stem Cell) whole cell lysate at 20 µg

    Lane 2: Mouse spleen tissue lysate at 20 µg

    Lane 3: Rat cerebellum tissue lysate at 20 µg

    Secondary

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

    Predicted band size: 86 kDa

    Observed band size: 100 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast cell line) cells labelling NSUN2/SAKI with ab259941 at 1/1000 (0.503 μg/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor®488) antibody at 1/1000 dilution (Green). Confocal image showing weak cytoplasmic and strong nuclear staining in NIH/3T3 cells. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor®594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941)

    Immunohistochemical analysis of paraffin-embedded mouse skin tissue labeling NSUN2/SAKI with ab259941 at 1/200 dilution (2.515 μg/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Nuclear staining on mouse skin (PMID: 16713953). The section was incubated with ab259941 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Flow Cytometry (Intracellular) - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling NSUN2/SAKI with ab259941 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941)

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue labeling NSUN2/SAKI with ab259941 at 1/200 dilution (2.515 μg/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Nuclear staining on human colon carcinoma (PMID: 16713953). The section was incubated with ab259941 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Flow Cytometry (Intracellular) - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized ES-D3 (mouse embryonic multipotent stem cell) cells labelling NSUN2/SAKI with ab259941 at 1/50 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941)

    Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling NSUN2/SAKI with ab259941 at 1/200 dilution (2.515 μg/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Nuclear staining on rat colon.The section was incubated with ab259941 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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