AB315016

Anti-53BP1 (phospho S25) antibody [EPR27057-33] - BSA and Azide free

RabMAb
KO Validated
Recombinant
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Knockout Tested Rabbit Recombinant Monoclonal 53BP1 phospho S25 antibody. Carrier free. Suitable for WB, Dot and reacts with Human, Synthetic peptide - Human samples.

View Alternative Names

TP53-binding protein 1, 53BP1, p53-binding protein 1, p53BP1, TP53BP1

4 Images

Supplier Data

Western blot - Anti-53BP1 (phospho S25) antibody [EPR27057-33] - BSA and Azide free (AB315016)

This data was developed using ab315015, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

The identity of the bands below 300 kDa is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

In Western blot, Anti-53BP1 antibody [EPR2172(2)] (ab175933) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-53BP1 (phospho S25) antibody [EPR27057-33] (<a href='/en-us/products/primary-antibodies/53bp1-phospho-s25-antibody-epr27057-33-ab315015'>ab315015</a>) at 1/1000 dilution

Lane 1:

Untreated HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

HeLa treated with 20J/m2 UV, then recovery for 1 hour, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 350 kDa,124 kDa

false

Exposure time: 10s

Supplier Data

Western blot - Anti-53BP1 (phospho S25) antibody [EPR27057-33] - BSA and Azide free (AB315016)

This data was developed using ab315015, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The identity of the bands below 300 kDa is unknown.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

In Western blot, Anti-53BP1 antibody [EPR2172(2)] (ab175933) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-53BP1 (phospho S25) antibody [EPR27057-33] (<a href='/en-us/products/primary-antibodies/53bp1-phospho-s25-antibody-epr27057-33-ab315015'>ab315015</a>) at 1/1000 dilution

Lane 1:

Untreated U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

U-2 OS treated with 10um Camptothecin for 1 hour, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 350 kDa,124 kDa

false

Exposure time: 3s

Supplier Data

Western blot - Anti-53BP1 (phospho S25) antibody [EPR27057-33] - BSA and Azide free (AB315016)

This data was developed using ab315015, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The identity of the bands below 300 kDa is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

In Western blot, Anti-53BP1 antibody [EPR2172(2)] (ab175933) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-53BP1 (phospho S25) antibody [EPR27057-33] (<a href='/en-us/products/primary-antibodies/53bp1-phospho-s25-antibody-epr27057-33-ab315015'>ab315015</a>) at 1/1000 dilution

Lane 1:

Untreated wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell line) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

Wild-type HAP1 treated with 100nM Calycin A for 30 minutes, whole cell lysate (untreated membrane) at 20 µg

Lane 3:

Untreated TP53BP1 knockout HAP1 whole cell lysate (untreated membrane) at 20 µg

Lane 4:

TP53BP1 knockout HAP1 treated with 100nM Calycin A for 30 minutes, whole cell lysate (untreated membrane) at 20 µg

Lane 5:

Untreated wild-type HAP1 whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 6:

Wild-type HAP1 treated with 100nM Calycin A for 30 minutes, whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 7:

Untreated TP53BP1 knockout HAP1 whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 8:

TP53BP1 knockout HAP1 treated with 100nM Calycin A for 30 minutes, whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 350 kDa,124 kDa

false

Exposure time: 3s

Supplier Data

Dot Blot - Anti-53BP1 (phospho S25) antibody [EPR27057-33] - BSA and Azide free (AB315016)

This data was developed using ab315015, the same antibody clone in a different buffer formulation.

Dot blot analysis of 53BP1 (phospho S25) using ab315015 at 1 : 1000 (0.521 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Exposure time : 180 seconds

Blocking and diluting buffer and concentration : 5% NFDM/TBST

The antibody recognizes 53BP1 diphosphorylated at S25 and S29. It does not cross-react with 53BP1 singly phosphorylated at S29.

Unconjugated

Anti-53BP1 (phospho S25) antibody [EPR27057-33]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR27057-33

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

Dot, WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Specificity

Non-specific bands at ~270 KDa and ~150 KDa have been observed by WB. The identity of these bands is unknown.

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Reactivity data

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Product details

ab315016 is the carrier-free version of ab315015.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

Compatibility
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.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

53BP1 also known as p53-binding protein 1 is a protein that plays an important role in DNA damage response. The 53BP1 protein has a molecular weight of approximately 213 kDa. It is a nuclear protein and it mainly expresses in cells during the DNA damage repair processes. Researchers commonly utilize immunofluorescence and anti-53BP1 antibodies to detect its presence especially for studying DNA repair mechanisms. 53BP1 proves important for maintenance of genomic stability.

Biological function summary

As a DNA damage response mediator 53BP1 functions to recognize and bind to sites of double-strand breaks in DNA promoting non-homologous end joining (NHEJ). This protein is a component of a larger protein complex that recruits factors necessary for DNA repair. Its interaction with DNA lesions also involves chromatin remodeling aiming to facilitate successful repair of the DNA. 53BP1 works in concert with other proteins such as H2AX and MDC1.

Pathways

Researchers observe 53BP1 in processes such as the DNA damage checkpoint and repair pathways. This protein significantly contributes to the pathways involved in maintaining cell cycle checkpoints. It interacts with proteins like ATM and BRCA1 coordinating the cellular response to DNA damage and ensuring that genomic integrity checks occur properly before cell cycle progression continues. These interactions place 53BP1 at a pivotal position in decision-making between repair pathways.

Researchers often focus on 53BP1's involvement in cancer development and therapy resistance due to its role in DNA repair. Alterations or deficiencies in 53BP1 levels can contribute to increased susceptibility to tumor development affecting processes like breast cancer and ovarian cancer through its relationship with BRCA1. The improper function of 53BP1 in DNA repair pathways may lead to an accumulation of genetic mutations driving tumorigenesis and impacting the efficacy of certain cancer treatments.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

Double-strand break (DSB) repair protein involved in response to DNA damage, telomere dynamics and class-switch recombination (CSR) during antibody genesis (PubMed : 12364621, PubMed : 17190600, PubMed : 21144835, PubMed : 22553214, PubMed : 23333306, PubMed : 27153538, PubMed : 28241136, PubMed : 31135337, PubMed : 37696958). Plays a key role in the repair of double-strand DNA breaks (DSBs) in response to DNA damage by promoting non-homologous end joining (NHEJ)-mediated repair of DSBs and specifically counteracting the function of the homologous recombination (HR) repair protein BRCA1 (PubMed : 22553214, PubMed : 23333306, PubMed : 23727112, PubMed : 27153538, PubMed : 31135337). In response to DSBs, phosphorylation by ATM promotes interaction with RIF1 and dissociation from NUDT16L1/TIRR, leading to recruitment to DSBs sites (PubMed : 28241136). Recruited to DSBs sites by recognizing and binding histone H2A monoubiquitinated at 'Lys-15' (H2AK15Ub) and histone H4 dimethylated at 'Lys-20' (H4K20me2), two histone marks that are present at DSBs sites (PubMed : 17190600, PubMed : 23760478, PubMed : 27153538, PubMed : 28241136). Required for immunoglobulin class-switch recombination (CSR) during antibody genesis, a process that involves the generation of DNA DSBs (PubMed : 23345425). Participates in the repair and the orientation of the broken DNA ends during CSR (By similarity). In contrast, it is not required for classic NHEJ and V(D)J recombination (By similarity). Promotes NHEJ of dysfunctional telomeres via interaction with PAXIP1 (PubMed : 23727112).

See full target information TP53BP1 phospho S25

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-53BP1 (phospho S25) antibody [EPR27057-33] - BSA and Azide free

Western blot - Anti-53BP1 (phospho S25) antibody [EPR27057-33] - BSA and Azide free (AB315016)

Western blot - Anti-53BP1 (phospho S25) antibody [EPR27057-33] - BSA and Azide free (AB315016)

Western blot - Anti-53BP1 (phospho S25) antibody [EPR27057-33] - BSA and Azide free (AB315016)

Dot Blot - Anti-53BP1 (phospho S25) antibody [EPR27057-33] - BSA and Azide free (AB315016)

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Specificity

Complementary Products

Primary Antibodies

Primary Antibodies

Secondary Antibodies

Primary Antibodies

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Product promise