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.
IgG
Rabbit
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
ICC/IF | IP | WB | Flow Cyt (Intra) | IHC-P | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Tested |
Mouse | Tested | Tested | Tested | Tested | Tested |
Rat | Expected | Expected | Tested | Expected | Tested |
Species | Dilution info | Notes |
---|---|---|
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. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
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 - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes - |
Species Rat | Dilution info 1/1000 | Notes - |
Species Human | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
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. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
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. |
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).
SAKI, TRM4, NSUN2, SAKI, TRM4, RNA cytosine C(5)-methyltransferase NSUN2, Myc-induced SUN domain-containing protein, NOL1/NOP2/Sun domain family member 2, Substrate of AIM1/Aurora kinase B, mRNA cytosine C(5)-methyltransferase, tRNA cytosine C(5)-methyltransferase, tRNA methyltransferase 4 homolog, Misu, hTrm4
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.
IgG
Rabbit
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR24140-93
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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:
For more information, read more on recombinant antibodies.
This supplementary information is collated from multiple sources and compiled automatically.
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.
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.
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.
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.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our 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.
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
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
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
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.
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.
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
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
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
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.
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.
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.
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.
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.
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.
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com