Anti-Nac1 antibody (ab29047)
Key features and details
- Rabbit polyclonal to Nac1
- Suitable for: IHC-P, WB, ICC/IF
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-Nac1 antibody
See all Nac1 primary antibodies -
Description
Rabbit polyclonal to Nac1 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 450 to the C-terminus of Mouse Nac1.
Read Abcam's proprietary immunogen policy (Peptide available as ab30604.) -
Positive control
- 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.
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
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.
Application Abreviews Notes 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. Target
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Function
Functions 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 specificity
Overexpressed 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 similarities
Contains 1 BEN domain.
Contains 1 BTB (POZ) domain. -
Cellular localization
Nucleus. Cytoplasm. Distribution in the cytoplasm is dependent on phosphorylation. - Information by UniProt
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Database links
- Entrez Gene: 112939 Human
- Entrez Gene: 66830 Mouse
- Entrez Gene: 171454 Rat
- Omim: 610672 Human
- SwissProt: Q96RE7 Human
- SwissProt: Q7TSZ8 Mouse
- SwissProt: O35260 Rat
- Unigene: 531614 Human
see all -
Alternative names
- BEND8 antibody
- BTB Domain Containing 14B antibody
- BTB/POZ domain-containing protein 14B antibody
see all
Images
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Western blot - Anti-Nac1 antibody (ab29047)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).
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Immunocytochemistry/ Immunofluorescence - Anti-Nac1 antibody (ab29047)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.
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Western blot - Anti-Nac1 antibody (ab29047)Anti-Nac1 antibody (ab29047) at 1/250 dilution + Brain (Rat) Tissue Lysate - normal tissue at 10 µg
Secondary
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 -
Immunocytochemistry/ Immunofluorescence - Anti-Nac1 antibody (ab29047)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). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nac1 antibody (ab29047)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.
Protocols
Datasheets and documents
References (6)
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
Images
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Western blot - Anti-Nac1 antibody (ab29047)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).
-
Immunocytochemistry/ Immunofluorescence - Anti-Nac1 antibody (ab29047)
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.
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Western blot - Anti-Nac1 antibody (ab29047)Anti-Nac1 antibody (ab29047) at 1/250 dilution + Brain (Rat) Tissue Lysate - normal tissue at 10 µg
Secondary
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
-
Immunocytochemistry/ Immunofluorescence - Anti-Nac1 antibody (ab29047)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).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nac1 antibody (ab29047)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.