Recombinant Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free (ab249845)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2173(2)] to 53BP1 - BSA and Azide free
- Suitable for: ICC/IF, WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-53BP1 antibody [EPR2173(2)] - BSA and Azide free
See all 53BP1 primary antibodies -
Description
Rabbit monoclonal [EPR2173(2)] to 53BP1 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WB, IHC-Pmore details
Unsuitable for: Flow Cyt or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1, HeLa, and HepG2 lysates. Human fetal brain and heart lysates. IHC-P: Human cervical carcinoma and prostate hyperplasia tissue. ICC/IF: HepG2 cells.
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General notes
ab249845 is the carrier-free version of ab175188.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Affinity purified -
Clonality
Monoclonal -
Clone number
EPR2173(2) -
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 ab249845 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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ICC/IF |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 214 kDa.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 214 kDa. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Function
May have a role in checkpoint signaling during mitosis. Enhances TP53-mediated transcriptional activation. Plays a role in the response to DNA damage. -
Involvement in disease
Note=A chromosomal aberration involving TP53BP1 is found in a form of myeloproliferative disorder chronic with eosinophilia. Translocation t(5;15)(q33;q22) with PDGFRB creating a TP53BP1-PDGFRB fusion protein. -
Sequence similarities
Contains 2 BRCT domains. -
Post-translational
modificationsAsymmetrically dimethylated on Arg residues by PRMT1. Methylation is required for DNA binding.
Phosphorylated at basal level in the absence of DNA damage. Hyper-phosphorylated in an ATM-dependent manner in response to DNA damage induced by ionizing radiation. Hyper-phosphorylated in an ATR-dependent manner in response to DNA damage induced by UV irradiation. -
Cellular localization
Nucleus. Chromosome > centromere > kinetochore. Associated with kinetochores. Both nuclear and cytoplasmic in some cells. Recruited to sites of DNA damage, such as double stand breaks. Methylation of histone H4 at 'Lys-20' is required for efficient localization to double strand breaks. - Information by UniProt
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Database links
- Entrez Gene: 7158 Human
- Omim: 605230 Human
- SwissProt: Q12888 Human
- Unigene: 440968 Human
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Alternative names
- 53 BP1 antibody
- 53BP1 antibody
- FLJ41424 antibody
see all
Images
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All lanes : Anti-53BP1 antibody [EPR2173(2)] (ab175188) at 1/50000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : TP53BP1 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 214 kDaThis data was developed using ab175188, the same antibody clone in a different buffer formulation.
Lanes 1 - 3: Merged signal (red and green). Green - ab175188 observed at 213 kDa. Red - loading control, ab130007, observed at 130 kDa.
ab175188 was shown to recognize 53BP1 in wild-type HAP1 cells as signal was lost at the expected MW in TP53BP1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and TP53BP1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab175188 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/50000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-53BP1 antibody [EPR2173(2)] (ab175188) at 1/50000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : TP53BP1 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 214 kDaThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175188).
Lanes 1 - 3: Merged signal (red and green). Green - ab175188 observed at 213 kDa. Red - loading control, ab130007, observed at 130 kDa.
ab175188 was shown to recognize 53BP1 in wild-type HAP1 cells as signal was lost at the expected MW in TP53BP1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and TP53BP1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab175188 and ab130007 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/50000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-53BP1 antibody [EPR2173(2)] (ab175188) at 1/50000 dilution
Lane 1 : Human fetal heart tissue lysate
Lane 2 : HepG2 cell lysate
Lane 3 : Human fetal brain tissue lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 214 kDaThis data was developed using ab175188, the same antibody clone in a different buffer formulation.
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This data was developed using ab175188, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue labeling 53BP1 with ab175188 at a 1/50 dilution. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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This data was developed using ab175188, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human prostate hyperplasia tissue labeling 53BP1 with ab175188 at a 1/50 dilution. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab249845 has not yet been referenced specifically in any publications.