Key features and details
- Rabbit polyclonal to Nac1
- Suitable for: IHC-P, WB, ICC/IF
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Product nameAnti-Nac1 antibody
DescriptionRabbit polyclonal to Nac1
Tested applicationsSuitable for: IHC-P, WB, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
- This antibody gave a positive signal in the following lysates: F9 (Mouse embryonic carcinoma cell line) Whole Cell IOUD2 (Mouse embryonic stem cell, selected for Oct4 expression cell line) Whole Cell Brain (Rat) Tissue Lysate - normal tissue It also gave a positive result in FFPE human cerebral cortex sections.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab29047 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||1/250. Detects a band of approximately 57,60 kDa (predicted molecular weight: 57 kDa).|
|ICC/IF||Use a concentration of 1 µg/ml.|
FunctionFunctions as a transcriptional repressor. Seems to function as a transcriptional corepressor in neuronal cells through recruitment of HDAC3 and HDAC4. Contributes to tumor progression, and tumor cell proliferation and survival. This may be mediated at least in part through repressing transcriptional activity of GADD45GIP1. Required for recruiting the proteasome from the nucleus to the cytoplasm and dendritic spines.
Tissue specificityOverexpressed in several types of carcinomas including ovarian serous carcinomas. Expression levels positively correlate with tumor recurrence in ovarian serous carcinomas, and intense immunoreactivity in primary ovarian tumors predicts early recurrence. Up-regulated in ovarian carcinomas after chemotherapy, suggesting a role in development of chemotherapy resistance in ovarian cancer.
Sequence similaritiesContains 1 BEN domain.
Contains 1 BTB (POZ) domain.
Cellular localizationNucleus. Cytoplasm. Distribution in the cytoplasm is dependent on phosphorylation.
- Information by UniProt
- BEND8 antibody
- BTB Domain Containing 14B antibody
- BTB/POZ domain-containing protein 14B antibody
We are unsure as to the nature of the doublet. It is likely that the doublet is caused by the presence of processed and unprocessed forms of Nac1. Both bands are blocked by addition of the immunizing peptide (ab30604).
ab29047 stained in Hela cells. Cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab29047 at 1µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Anti-Nac1 antibody (ab29047) at 1/250 dilution + Brain (Rat) Tissue Lysate - normal tissue at 10 µg
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 57 kDa
Observed band size: 57 kDa
ICC/IF image of ab29047 stained mouse embryonic stem cells. The cells were 4% PFA 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 (ab29047, 1µg/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).
IHC image of Nac1 staining in human cerebral cortex 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 ab29047, 5µg/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.
ab29047 has been referenced in 6 publications.
- Ju T et al. Overexpression of NAC1 confers drug resistance via HOXA9 in colorectal carcinoma cells. Mol Med Rep 16:3194-3200 (2017). PubMed: 28713930
- Han H et al. Multilayered Control of Alternative Splicing Regulatory Networks by Transcription Factors. Mol Cell 65:539-553.e7 (2017). PubMed: 28157508
- Malleshaiah M et al. Nac1 Coordinates a Sub-network of Pluripotency Factors to Regulate Embryonic Stem Cell Differentiation. Cell Rep 14:1181-1194 (2016). PubMed: 26832399
- Sloan E et al. Analysis of the SUMO2 Proteome during HSV-1 Infection. PLoS Pathog 11:e1005059 (2015). PubMed: 26200910
- Gagliardi A et al. A direct physical interaction between Nanog and Sox2 regulates embryonic stem cell self-renewal. EMBO J 32:2231-47 (2013). WB . PubMed: 23892456
- MacArthur BD et al. Nanog-dependent feedback loops regulate murine embryonic stem cell heterogeneity. Nat Cell Biol 14:1139-47 (2012). WB ; Mouse . PubMed: 23103910