Recombinant Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)
![Western blot - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)](https://www.abcam.com/ps/products/196/ab196022/Images/ab196022-261791-anti-baf180-antibody-epr15860-western-blot.jpg "Western blot - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR15860] to PBRM1/BAF180
- Suitable for: Flow Cyt (Intra), IHC-P, WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-PBRM1/BAF180 antibody [EPR15860]
See all PBRM1/BAF180 primary antibodies -
Description
Rabbit monoclonal [EPR15860] to PBRM1/BAF180 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat
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Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human fetal kidney lysate; HeLa nuclear lysate; IHC-P: Human kidney tissue; ICC/IF: 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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR15860 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab196022 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 |
|---|---|---|
| Flow Cyt (Intra) |
Use at an assay dependent concentration.
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| IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| WB |
1/10000. Detects a band of approximately 193 kDa (predicted molecular weight: 193 kDa).
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| ICC/IF |
1/100.
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| Notes |
|---|
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
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IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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WB
1/10000. Detects a band of approximately 193 kDa (predicted molecular weight: 193 kDa). |
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ICC/IF
1/100. |
Target
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Function
Involved in transcriptional activation and repression of select genes by chromatin remodeling (alteration of DNA-nucleosome topology). Act as a negative regulator of cell proliferation. -
Tissue specificity
Widely expressed. -
Involvement in disease
Defects in PBRM1 are a cause of renal cell carcinoma (RCC) [MIM:144700]. It is a heterogeneous group of sporadic or hereditary carcinoma derived from cells of the proximal renal tubular epithelium. It is subclassified into clear cell renal carcinoma (non-papillary carcinoma), papillary renal cell carcinoma, chromophobe renal cell carcinoma, collecting duct carcinoma with medullary carcinoma of the kidney, and unclassified renal cell carcinoma. -
Sequence similarities
Contains 2 BAH domains.
Contains 6 bromo domains.
Contains 1 HMG box DNA-binding domain. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 55193 Human
- Entrez Gene: 66923 Mouse
- Entrez Gene: 306254 Rat
- Omim: 606083 Human
- SwissProt: Q86U86 Human
- SwissProt: Q8BSQ9 Mouse
- Unigene: 189920 Human
- Unigene: 476320 Human
see all -
Alternative names
- BAF180 antibody
- BRG1-associated factor 180 antibody
- CG11375 antibody
see all
Images
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Western blot - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: PBRM1/BAF180 knockout HAP1 cell lysate (20 µg)
Lane 3: Human kidney tissue lysate (20 µg)
Lane 4: HeLa cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab196022 observed at 193 kDa. Red - loading control, ab18058, observed at 124 kDa.
ab196022 was shown to recognize PBRM1/BAF180 when PBRM1/BAF180 knockout samples were used, along with additional cross-reactive bands. Wild-type and PBRM1/BAF180 knockout samples were subjected to SDS-PAGE. ab196022 and ab18058 (loading control to Vinculin) were diluted 1/5000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging. -
![Flow Cytometry (Intracellular) - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)](https://www.abcam.com/ps/products/196/ab196022/Images/ab196022-263930-anti-baf180-antibody-epr15860-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)ab196022 staining PBRM1/BAF180 in the human cell line HeLa (human cervix adenocarcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. The sample was incubated with the primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
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![Western blot - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)](https://www.abcam.com/ps/products/196/ab196022/Images/ab196022-241782-anti-baf180-antibody-epr15860-western-blot.jpg)
Western blot - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022) at 1/10000 dilution + Human fetal kidney lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 193 kDa
Observed band size: 193,35 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutes5% NFDM/TBST: Blocking and diluting buffer.
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![Western blot - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)](https://www.abcam.com/ps/products/196/ab196022/Images/ab196022-241781-anti-baf180-antibody-epr15860-western-blot.jpg)
Western blot - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022) at 1/10000 dilution + HeLa (Human epithelial cells from cervix adenocarcinoma) nuclear lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 193 kDa
Observed band size: 193 kDa
Exposure time: 15 seconds5% NFDM/TBST: Blocking and diluting buffer.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)](https://www.abcam.com/ps/products/196/ab196022/Images/ab196022-241779-anti-baf180-antibody-epr15860-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling PBRM1/BAF180 using ab196022 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human kidney tissue is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Immunocytochemistry/ Immunofluorescence - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)](https://www.abcam.com/ps/products/196/ab196022/Images/ab196022-241778-anti-baf180-antibody-epr15860-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PBRM1/BAF180 with ab196022 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). The nuclear counter stain is DAPI (blue).
Nuclear staining on HeLa cell line is observed.
-ve control - Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution. The nuclear counter stain is DAPI (blue).
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![Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)](https://www.abcam.com/ps/products/196/ab196022/Images/ab196022-7-benefits-of-recombinant-antibodies.png)
Anti-PBRM1/BAF180 antibody [EPR15860] (ab196022)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (3)
ab196022 has been referenced in 3 publications.
- Cai W et al. Expression of PBRM1 as a prognostic predictor in metastatic renal cell carcinoma patients treated with tyrosine kinase inhibitor. Int J Clin Oncol 25:338-346 (2020). PubMed: 31720994
- Liu T et al. CCL5-dependent mast cell infiltration into the tumor microenvironment in clear cell renal cell carcinoma patients. Aging (Albany NY) 12:21809-21836 (2020). PubMed: 33177244
- Cabot B et al. Differential expression of key subunits of SWI/SNF chromatin remodeling complexes in porcine embryos derived in vitro or in vivo. Mol Reprod Dev 84:1238-1249 (2017). ICC/IF . PubMed: 29024220