Recombinant Anti-BANF1/BAF antibody [EPR7668] (ab129184)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7668] to BANF1/BAF
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Reacts with: Mouse, Dog, Human
Related conjugates and formulations
Overview
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Product name
Anti-BANF1/BAF antibody [EPR7668]
See all BANF1/BAF primary antibodies -
Description
Rabbit monoclonal [EPR7668] to BANF1/BAF -
Host species
Rabbit -
Specificity
The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse.
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Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Dog, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, HepG2, MDCK, and HEK-293T cell lysates. IHC-P: Human colonic carcinoma, and ovarian carcinoma tissues. ICC/IF: HeLa, HepG2 and NIH/3T3 cells. Flow Cyt (intra): HeLa cells.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles. -
Dissociation constant (KD)
KD = 2.18 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR7668 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab129184 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 |
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Flow Cyt (Intra) |
1/80.
For unpurified use at 1/200 |
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WB | (1) |
1/1000 - 1/10000. Predicted molecular weight: 10 kDa.
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IHC-P | (2) |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF | (2) |
1/100.
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Notes |
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Flow Cyt (Intra)
1/80. For unpurified use at 1/200 |
WB
1/1000 - 1/10000. Predicted molecular weight: 10 kDa. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/100. |
Target
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Function
Plays fundamental roles in nuclear assembly, chromatin organization, gene expression and gonad development. May potently compress chromatin structure and be involved in membrane recruitment and chromatin decondensation during nuclear assembly. Contains 2 non-specific dsDNA-binding sites which may promote DNA cross-bridging. Exploited by retroviruses for inhibiting self-destructing autointegration of retroviral DNA, thereby promoting integration of viral DNA into the host chromosome. EMD and BAF are cooperative cofactors of HIV-1 infection. Association of EMD with the viral DNA requires the presence of BAF and viral integrase. The association of viral DNA with chromatin requires the presence of BAF and EMD. -
Tissue specificity
Widely expressed. Expressed in colon, brain, heart, kidney, liver, lung, ovary, pancreas, placenta, prostate, skeletal muscle, small intestine, spleen and testis. Not detected in thymus and peripheral blood leukocytes. -
Sequence similarities
Belongs to the BAF family. -
Domain
Has a helix-hairpin-helix (HhH) structural motif conserved among proteins that bind non-specifically to DNA.
LEM domain proteins bind centrally on the BAF dimer, whereas DNA binds to the left and right sides. -
Post-translational
modificationsPartially phosphorylated on serine. Ser-4 phosphorylation may block BAF ability to promote EMD binding to lamins in vitro. Non phosphorylated BAF seems to enhances binding between EMD and LMNA. -
Cellular localization
Nucleus. Cytoplasm. Chromosome. Significantly enriched at the nuclear inner membrane, diffusely throughout the nucleus during interphase and concentrated at the chromosomes during the M-phase. May be included in HIV-1 virions via its interaction with viral GAG polyprotein. - Information by UniProt
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Database links
- Entrez Gene: 8815 Human
- Entrez Gene: 23825 Mouse
- Omim: 603811 Human
- SwissProt: O75531 Human
- SwissProt: O54962 Mouse
- Unigene: 433759 Human
- Unigene: 358649 Mouse
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Alternative names
- BAF antibody
- BAF_HUMAN antibody
- BANF 1 antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human ovarian carcinoma tissue sections labeling BANF1/BAF with Purified ab129184 at 1:100 dilution (0.78 µg/ml). Heat mediated antigen retrieval was performed using Citrate buffer, pH 6.0. ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling BANF1/BAF with Purified ab129184 at 1:100 dilution (7.8 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Anti-BANF1/BAF antibody [EPR7668] (ab129184) at 1/5000 dilution + MDCK (NBL-2) (Dog kidney epithelial cell) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 10 kDa
Observed band size: 10 kDaBlocking Buffer and concentration: 5% NFDM/TBST
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All lanes : Anti-BANF1/BAF antibody [EPR7668] (ab129184) at 1/1000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 10 kDa
Observed band size: 10 kDaBlocking Buffer and concentration: 5% NFDM/TBST
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All lanes : Anti-BANF1/BAF antibody [EPR7668] (ab129184) at 1/1000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysates
Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) cell lysates
Lane 3 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 10 kDa -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryo fibroblast cells)cells labeling BANF1/BAF with ab129184 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing mainly nuclear with cytoplasmic staining on NIH/3T3 cell line. The nuclear counterstain is DAPI (blue).
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ab129184 staining BANF1/BAF in MDCK (Canine kidney cell line) cell pellets in paraffin by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Cell pellets were fixed with paraformaldehyde, permeabilized with Tween-20 and blocked with 1% serum for 2 hours at room temperature; antigen retrieval was by heat mediation in Tris/EDTA pH 9. Samples were incubated with primary antibody (1/100 in 1% BSA + 1% FBS) for 16 hours. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.
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Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling BANF1/BAF (red) with ab129184 at a 1/200 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with primary and secondary antibodies.
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ab129184, at a 1/100 dilution, staining BANF1/BAF in paraffin embedded Human colonic carcinoma tissue by Immunohistochemistry.
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma) cells labeling BANF1/BAF with ab129184 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing mainly nuclear with weakly cytoplasmic staining on Hela cell line. The nuclear counterstain is DAPI (blue).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab129184 has been referenced in 10 publications.
- Sears RM & Roux KJ Mechanisms of A-Type Lamin Targeting to Nuclear Ruptures Are Disrupted in LMNA- and BANF1-Associated Progerias. Cells 11:N/A (2022). PubMed: 35269487
- Prissette M et al. Disruption of nuclear envelope integrity as a possible initiating event in tauopathies. Cell Rep 40:111249 (2022). PubMed: 36001963
- Janssen A et al. The BAF A12T mutation disrupts lamin A/C interaction, impairing robust repair of nuclear envelope ruptures in Nestor-Guillermo progeria syndrome cells. Nucleic Acids Res 50:9260-9278 (2022). PubMed: 36039758
- Kono Y et al. Nucleoplasmic lamin C rapidly accumulates at sites of nuclear envelope rupture with BAF and cGAS. J Cell Biol 221:N/A (2022). IHC-P ; Human . PubMed: 36301259
- Mohr L et al. ER-directed TREX1 limits cGAS activation at micronuclei. Mol Cell 81:724-738.e9 (2021). PubMed: 33476576
- Essawy N et al. An Emerin LEM-Domain Mutation Impairs Cell Response to Mechanical Stress. Cells 8:N/A (2019). PubMed: 31185657
- Halfmann CT et al. Repair of nuclear ruptures requires barrier-to-autointegration factor. J Cell Biol 218:2136-2149 (2019). PubMed: 31147383
- Elkhatib RA et al. LEM-domain proteins are lost during human spermiogenesis but BAF and BAF-L persist. Reproduction 154:387-401 (2017). PubMed: 28684548
- Birendra Kc et al. VRK2A is an A-type lamin-dependent nuclear envelope kinase that phosphorylates BAF. Mol Biol Cell 28:2241-2250 (2017). PubMed: 28637768
- Paci M et al. Abnormal retention of nuclear lamina and disorganization of chromatin-related proteins in spermatozoa from DPY19L2-deleted globozoospermic patients. Reprod Biomed Online 35:562-570 (2017). PubMed: 28882431