Key features and details
- Rabbit polyclonal to nmt55 / p54nrb
- Suitable for: ICC/IF, IHC-P, WB, IP
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Product nameAnti-nmt55 / p54nrb antibody
See all nmt55 / p54nrb primary antibodies
DescriptionRabbit polyclonal to nmt55 / p54nrb
Tested applicationsSuitable for: ICC/IF, IHC-P, WB, IPmore details
Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Rabbit, Horse, Guinea pig, Cow, Dog, Pig, Chimpanzee, Rhesus monkey, Gorilla, Orangutan, Elephant
Synthetic peptide corresponding to a region between residues 350 and 400 of human nmt55/ p54nrb (NP_031389.3).
- Whole cell lysates from HeLa and 293T cells.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 6.8
Preservative: 0.09% Sodium azide
Constituents: 0.1% BSA, Tris buffered saline
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab70335 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|WB||1/2000 - 1/10000. Detects a band of approximately 60 kDa (predicted molecular weight: 54 kDa).|
|IP||Use at 1-4 µg/mg of lysate.|
FunctionDNA- and RNA binding protein, involved in several nuclear processes. Binds the conventional octamer sequence in double stranded DNA. Also binds single-stranded DNA and RNA at a site independent of the duplex site (By similarity). Involved in pre-mRNA splicing, probably as an heterodimer with SFPQ. Interacts with U5 snRNA, probably by binding to a purine-rich sequence located on the 3' side of U5 snRNA stem 1b. The SFPQ-NONO heteromer associated with MATR3 may play a role in nuclear retention of defective RNAs. The SFPQ-NONO heteromer may be involved in DNA unwinding by modulating the function of topoisomerase I/TOP1. The SFPQ-NONO heteromer may be involved in DNA nonhomologous end joining (NHEJ) required for double-strand break repair and V(D)J recombination and may stabilize paired DNA ends. In vitro, the complex strongly stimulates DNA end joining, binds directly to the DNA substrates and cooperates with the Ku70/G22P1-Ku80/XRCC5 (Ku) dimer to establish a functional preligation complex. Nono is involved in transcriptional regulation. The SFPQ-NONO-NR5A1 complex binds to the CYP17 promoter and regulates basal and cAMP-dependent transcriptional avtivity. NONO binds to an enhancer element in long terminal repeats of endogenous intracisternal A particles (IAPs) and activates transcription.
Tissue specificityHeart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas. Also found in a number of breast tumor cell lines.
Involvement in diseaseNote=A chromosomal aberration involving NONO may be a cause of papillary renal cell carcinoma (PRCC). Translocation t(X;X)(p11.2;q13.1) with TFE3.
Sequence similaritiesContains 2 RRM (RNA recognition motif) domains.
modificationsThe N-terminus is blocked.
- Information by UniProt
- 52 kDa subunit antibody
- 54 kDa nuclear RNA and DNA binding protein antibody
- 54 kDa nuclear RNA- and DNA-binding protein antibody
All lanes : Anti-nmt55 / p54nrb antibody (ab70335) at 0.04 µg/ml
Lane 1 : Whole cell lysate from HeLa cells at 50 µg
Lane 2 : Whole cell lysate from HeLa cells at 15 µg
Lane 3 : Whole cell lysate from HeLa cells at 5 µg
Lane 4 : Whole cell lysate from 293T cells at 50 µg
Predicted band size: 54 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma (left) and mouse squamous cell carcinoma (right) tissues labelling nmt55 / p54nrb with ab70335 at 1/1000 (0.2µg/ml) and 1/200 (1µg/ml). Detection: DAB.
Detection of Human nmt55 / p54nrb by Immunoprecipitation in Whole cell lysate from HeLa cells (1 mg for IP, 1/4 of IP loaded) using ab70335 at 3 µg/mg lysate for IP (Lane 1) and at 1 µg/ml for subsequent WB detection. Lane 2 represents control IgG IP.
ICC/IF image of ab70335 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab42905, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
IHC image of ab70335 staining in human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab70335, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
All lanes : Anti-nmt55 / p54nrb antibody (ab70335) at 0.1 µg/ml
Lane 1 : NIH3T3 whole cell lysate
Lane 2 : TCMK-1 whole cell lysate
Lane 3 : 4T1 whole cell lysate
Lane 4 : CT26.WT whole cell lysate
Lane 5 : C6 whole cell lysate
Lysates/proteins at 50 µg per lane.
Developed using the ECL technique.
Predicted band size: 54 kDa
Exposure time: 30 seconds
ab70335 has been referenced in 13 publications.
- Bajc Cesnik A et al. Nuclear RNA foci from C9ORF72 expansion mutation form paraspeckle-like bodies. J Cell Sci 132:N/A (2019). PubMed: 30745340
- Modic M et al. Cross-Regulation between TDP-43 and Paraspeckles Promotes Pluripotency-Differentiation Transition. Mol Cell 74:951-965.e13 (2019). PubMed: 31047794
- Yu JSE et al. The FUS-DDIT3 Interactome in Myxoid Liposarcoma. Neoplasia 21:740-751 (2019). PubMed: 31220736
- Vickers TA et al. Kinetic and subcellular analysis of PS-ASO/protein interactions with P54nrb and RNase H1. Nucleic Acids Res 47:10865-10880 (2019). PubMed: 31495875
- Li D et al. Ets-1 promoter-associated noncoding RNA regulates the NONO/ERG/Ets-1 axis to drive gastric cancer progression. Oncogene 37:4871-4886 (2018). PubMed: 29773901