Name: Anti-53BP1 antibody [EPR2172(2)]
SKU: ab175933
Rating: 5 (3 reviews)

Anti-53BP1 antibody [EPR2172(2)] (ab175933) is a rabbit monoclonal antibody that is used to detect 53BP1 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat samples.

- Specificity confirmed with 53BP1 knockout cell line validation

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Images

Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2172(2)] (AB175933), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

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

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	WB	ICC/IF	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested
Mouse	Tested	Tested	Expected	Expected
Rat	Tested	Tested	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/60 - 1/250	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.
Species Rat	Dilution info 1/60 - 1/250	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.
Species Human	Dilution info 1/60 - 1/250	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000 - 1/5000	Notes -
Species Rat	Dilution info 1/1000 - 1/5000	Notes -
Species Human	Dilution info 1/1000 - 1/5000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/250	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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7 products for Alternative Product

Target data

See full target information TP53BP1

Function

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).

Alternative names

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

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR2172(2)
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

23 product images

Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling 53BP1 with purified ab175933 at 1/200. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/200) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).
Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
Lane 1: Wild-type HAP1 cell lysate (20 μg)
Lane 2: 53BP1 knockout HAP1 cell lysate (20 μg)
Lane 3: HeLa cell lysate (40 μg)
Lane 4: HepG2 cell lysate (40 μg)
Lanes 1 - 4: Merged signal (red and green). Green - ab175933 observed at 350 kDa. Red - loading control, ab18058, observed at 124 kDa.
ab175933 was shown to specifically react with 53BP1 when 53BP1 knockout samples were used. Wild-type and 53BP1 knockout samples were subjected to SDS-PAGE. ab175933 and ab18058 (loading control to Vinculin) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
Predicted band size: 214 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue labelling 53BP1 with purified ab175933 at a dilution of 1/60. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Flow Cytometry (Intracellular) - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
ab175933 staining 53BP1in the human cell lineHepG2 (human hepatocellular carcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/30. 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)
Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
Lane 1: Wild-type HAP1 cell lysate (20 μg)
Lane 2: 53BP1 knockout HAP1 cell lysate (20 μg)
Lane 3: HeLa cell lysate (40 μg)
Lane 4: HepG2 cell lysate (40 μg)
Lanes 1 - 4: Merged signal (red and green).
Green - Target observed at 350 kDa. Red - loading control, ab18058, observed at 124 kDa.
This western blot image is a comparison between ab175933 and a competitor's top cited rabbit polyclonal antibody.
All lanes: Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
Predicted band size: 214 kDa
Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
Blocking and dilution buffer: 5% NFDM /TBST.
All lanes: Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933) at 1/1000 dilution
Lane 1: Mouse heart tissue lysate at 10 µg
Lane 2: Mouse brain tissue lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 214 kDa
Observed band size: 450 kDa
Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
Blocking and dilution buffer: 5% NFDM /TBST.
All lanes: Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933) at 20 µg
All lanes: Rat heart tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 214 kDa
Observed band size: 450 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue labelling 53BP1 with purified ab175933 at a dilution of 1/60. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
Blocking and dilution buffer: 5% NFDM /TBST.
All lanes: Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933) at 1/5000 dilution
Lane 1: Human fetal heart tissue lysate at 20 µg
Lane 2: HeLa whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 214 kDa
Observed band size: 450 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver carcinoma tissue labelling 53BP1 with purified ab175933 at a dilution of 1/60. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
All lanes: Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933) at 1/1000 dilution
Lane 1: HepG2 cell lysate at 1/10 dilution
Lane 2: Human fetal brain lysate at 1/10 dilution
Lane 3: Human fetal heart lysate at 1/10 dilution
Predicted band size: 214 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling 53BP1 with unpurified ab175933 at a dilution of 1/100.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling 53BP1 with unpurified ab175933 at a dilution of 1/100.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling 53BP1 with unpurified ab175933 at a dilution of 1/100.
Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
53BP1 Western blot staining using rabbit Anti-53BP1 antibody
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] (Anti-Vinculin antibody [EPR8185] - Loading Control 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] (Anti-53BP1 (phospho S25) antibody [EPR27057-33] ab315015) 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 350 kDa, 124 kDa
Exposure time: 3s
Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
53BP1 Western blot staining using rabbit Anti-53BP1 antibody
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] (Anti-Vinculin antibody [EPR8185] - Loading Control 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] (Anti-53BP1 (phospho S25) antibody [EPR27057-33] ab315015) 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 350 kDa, 124 kDa
Exposure time: 10s
Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
53BP1 Western blot staining using rabbit Anti-53BP1 antibody
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] (Anti-Vinculin antibody [EPR8185] - Loading Control 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] (Anti-53BP1 (phospho S25) antibody [EPR27057-33] ab315015) 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 350 kDa, 124 kDa
Exposure time: 3s
Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
53BP1 Western blot staining using rabbit Anti-53BP1 antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.
In Western blot, Anti-53BP1 antibody [EPR2172(2)] (ab175933) staining at 350kDa dilution.
All lanes: Western blot - Anti-53BP1 (phospho T670) antibody [EPR27060-30] (Anti-53BP1 (phospho T670) antibody [EPR27060-30] ab314866) at 1/1000 dilution
Lane 1: Untreated HeLa (human cervix adenocarcinoma epithelial cell) 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Developed using the ECL technique.
Observed band size: 350 kDa, 124 kDa
Exposure time: 114s
Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
53BP1 Western blot staining using rabbit Anti-53BP1 antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.
In Western blot, Anti-53BP1 antibody [EPR2172(2)] (ab175933) staining at 350kDa dilution.
All lanes: Western blot - Anti-53BP1 (phospho T670) antibody [EPR27060-30] (Anti-53BP1 (phospho T670) antibody [EPR27060-30] ab314866) at 1/1000 dilution
Lane 1: HeLa (human epithelial cell line from cervical adenocarcinoma) whole cell lysate at 20 µg
Lane 2: HeLa treated with 100ng/ml Calyculin A for 10 minutes, whole cell lysate at 20 µg
Lane 3: A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 4: A431 treated with 100ng/ml Calyculin A for 15 minutes, whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 350 kDa, 124 kDa
Exposure time: 180s
Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
53BP1 Western blot staining using rabbit Anti-53BP1 antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.
In Western blot, Anti-53BP1 antibody [EPR2172(2)] (ab175933) staining at 350kDa dilution.
All lanes: Western blot - Anti-53BP1 (phospho T670) antibody [EPR27060-30] (Anti-53BP1 (phospho T670) antibody [EPR27060-30] ab314866) 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(human chronic myelogenous leukemia near-haploid cell line) 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 350 kDa, 124 kDa
Exposure time: 180s
Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
53BP1 Western blot staining using rabbit Anti-53BP1 antibody
The identity of the band at 130 kDa is unknown.
The bands beneath the target band (350 kDa) are likely to be degraded target fragments .
In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control 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 T543) antibody [EPR27059-168] (Anti-53BP1 (phospho T543) antibody [EPR27059-168] ab316316) at 1/1000 dilution
Lane 1: Untreated HeLa (human epithelial cell line from cervical adenocarcinoma) whole cell lysate at 20 µg with 5% NFDM/TBST
Lane 2: HeLa treated with /ml Calyculin A for 15 minutes, whole cell lysate at 20 µg with 5% NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 350 kDa, 124 kDa
Exposure time: 180s
Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
53BP1 Western blot staining using rabbit Anti-53BP1 antibody
The identity of the band at 130 kDa is unknown.
The bands beneath the target band (350 kDa) are likely to be degraded target fragments which are with total protein control ab175933 .
In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control 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 T543) antibody [EPR27059-168] (Anti-53BP1 (phospho T543) antibody [EPR27059-168] ab316316) at 1/1000 dilution
Lane 1: Untreated U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg with 5% NFDM/TBST
Lane 2: U-2 OS treated with 10um Camptothecin for 1 hour, whole cell lysate (untreated membrane) at 20 µg with 5% NFDM/TBST
Lane 3: Untreated U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate (alkaline phosphatase treated membrane) at 20 µg with 5% NFDM/TBST
Lane 4: U-2 OS treated with 10um Camptothecin for 1 hour, whole cell lysate (alkaline phosphatase treated membrane) at 20 µg with 5% NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 350 kDa, 124 kDa
Exposure time: 48s
Western blot - Anti-53BP1 antibody [EPR2172(2)] (ab175933)
53BP1 Western blot staining using rabbit Anti-53BP1 antibody
The identity of the band at 130 kDa is unknown.
The bands beneath the target band (350 kDa) are likely to be degraded target fragments which are with total protein control ab175933.
In Western blot, Anti-53BP1 (phospho T543) antibody [EPR27059-168] ab316316 was shown to bind specifically to 53BP1 (phospho T543). A band was observed at 350 kDa in Calyculin A treated wild-type HAP1 cell lysates whereas loss of signal was observed in the 53BP1 knockout cell line.
In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control 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 T543) antibody [EPR27059-168] (Anti-53BP1 (phospho T543) antibody [EPR27059-168] ab316316) 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 with 5% NFDM/TBST
Lane 2: Wild-type HAP1 treated with 100nM Calycin A for 30 minutes, whole cell lysate (untreated membrane) at 20 µg with 5% NFDM/TBST
Lane 3: Untreated TP53BP1 knockout HAP1 whole cell lysate (untreated membrane) at 20 µg with 5% NFDM/TBST
Lane 4: TP53BP1 knockout HAP1 treated with 100nM Calycin A for 30 minutes, whole cell lysate (untreated membrane) at 20 µg with 5% NFDM/TBST
Lane 5: Untreated wild-type HAP1(human chronic myelogenous leukemia near-haploid cell line) whole cell lysate (alkaline phosphatase treated membrane) at 20 µg with 5% NFDM/TBST
Lane 6: Wild-type HAP1 treated with 100nM Calycin A for 30 minutes, whole cell lysate (alkaline phosphatase treated membrane) at 20 µg with 5% NFDM/TBST
Lane 7: Untreated TP53BP1 knockout HAP1 whole cell lysate (alkaline phosphatase treated membrane) at 20 µg with 5% NFDM/TBST
Lane 8: TP53BP1 knockout HAP1 treated with 100nM Calycin A for 30 minutes, whole cell lysate (alkaline phosphatase treated membrane) at 20 µg with 5% NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 350 kDa, 124 kDa
Exposure time: 48s