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Mouse Monoclonal P53 antibody. Carrier free. Suitable for ICC/IF, ChIP, Flow Cyt (Intra), WB, IHC-P and reacts with Human samples.

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Images

ChIP - Anti-p53 antibody [DO-1] - BSA and Azide free (AB237976), expandable thumbnail
  • Western blot - Anti-p53 antibody [DO-1] - BSA and Azide free (AB237976), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [DO-1] - BSA and Azide free (AB237976), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody [DO-1] - BSA and Azide free (AB237976), expandable thumbnail
  • Western blot - Anti-p53 antibody [DO-1] - BSA and Azide free (AB237976), expandable thumbnail

Key facts

Isotype

IgG2a

Host species

Mouse

Storage buffer

Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ICC/IFChIPFlow Cyt (Intra)WBIHC-P
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1-5 µg/mL

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Rat

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

Select an associated product type

1 product for Alternative Version

Target data

Function

Multifunctional transcription factor that induces cell cycle arrest, DNA repair or apoptosis upon binding to its target DNA sequence (PubMed:11025664, PubMed:12524540, PubMed:12810724, PubMed:15186775, PubMed:15340061, PubMed:17317671, PubMed:17349958, PubMed:19556538, PubMed:20673990, PubMed:20959462, PubMed:22726440, PubMed:24051492, PubMed:24652652, PubMed:35618207, PubMed:36634798, PubMed:38653238, PubMed:9840937). Acts as a tumor suppressor in many tumor types; induces growth arrest or apoptosis depending on the physiological circumstances and cell type (PubMed:11025664, PubMed:12524540, PubMed:12810724, PubMed:15186775, PubMed:15340061, PubMed:17189187, PubMed:17317671, PubMed:17349958, PubMed:19556538, PubMed:20673990, PubMed:20959462, PubMed:22726440, PubMed:24051492, PubMed:24652652, PubMed:38653238, PubMed:9840937). Negatively regulates cell division by controlling expression of a set of genes required for this process (PubMed:11025664, PubMed:12524540, PubMed:12810724, PubMed:15186775, PubMed:15340061, PubMed:17317671, PubMed:17349958, PubMed:19556538, PubMed:20673990, PubMed:20959462, PubMed:22726440, PubMed:24051492, PubMed:24652652, PubMed:9840937). One of the activated genes is an inhibitor of cyclin-dependent kinases. Apoptosis induction seems to be mediated either by stimulation of BAX and FAS antigen expression, or by repression of Bcl-2 expression (PubMed:12524540, PubMed:17189187). Its pro-apoptotic activity is activated via its interaction with PPP1R13B/ASPP1 or TP53BP2/ASPP2 (PubMed:12524540). However, this activity is inhibited when the interaction with PPP1R13B/ASPP1 or TP53BP2/ASPP2 is displaced by PPP1R13L/iASPP (PubMed:12524540). In cooperation with mitochondrial PPIF is involved in activating oxidative stress-induced necrosis; the function is largely independent of transcription. Induces the transcription of long intergenic non-coding RNA p21 (lincRNA-p21) and lincRNA-Mkln1. LincRNA-p21 participates in TP53-dependent transcriptional repression leading to apoptosis and seems to have an effect on cell-cycle regulation. Implicated in Notch signaling cross-over. Prevents CDK7 kinase activity when associated to CAK complex in response to DNA damage, thus stopping cell cycle progression. Isoform 2 enhances the transactivation activity of isoform 1 from some but not all TP53-inducible promoters. Isoform 4 suppresses transactivation activity and impairs growth suppression mediated by isoform 1. Isoform 7 inhibits isoform 1-mediated apoptosis. Regulates the circadian clock by repressing CLOCK-BMAL1-mediated transcriptional activation of PER2 (PubMed:24051492).

Alternative names

Recommended products

Mouse Monoclonal P53 antibody. Carrier free. Suitable for ICC/IF, ChIP, Flow Cyt (Intra), WB, IHC-P and reacts with Human samples.

Key facts

Isotype

IgG2a

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

DO-1

Purification technique

Affinity purification Protein G

Specificity

This antibody clone recognises both wild-type and mutant forms of p53 in human samples. It is not designed to recognise any specific p53 mutation.

We have confirmed this experimentally and have been able to detect p53 in different cell lines using various applications and treatments.

Important note: p53 expression levels vary greatly between cell lines. It has been reported that p53 mutations render the protein more stable, hence mutated cell lines often express higher levels of the p53 protein compared to wild-type cell lines. For low expressing wild type cell lines, p53 expression can be increased with cell treatments such as camptothecin or irinotecan.

Epitope

The epitope maps to within aa 20-25.

Light chain type

kappa

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab237976 is the carrier-free version of Anti-p53 antibody [DO-1] - ChIP Grade ab1101.

Abcam is leading the way to address reproducibility in scientific research with our highly validated recombinant monoclonal and recombinant multiclonal antibodies. Search & select one of Abcam's thousands of recombinant alternatives to eliminate batch-variability and unnecessary animal use.

If you do not find a host species to meet your needs, our catalogue and custom Chimeric range provides scientists the specificity of Abcam's RabMAbs in the species backbone of your choice. Remember to also review our range of edited cell lines, proteins and biochemicals relevant to your target that may help you further your research goals.

Abcam antibodies are extensively validated in a wide range of species and applications, so please check the reagent specifications meet your scientific needs before purchasing. If you have any questions or bespoke requirements, simply visit the Contact Us page to send us an inquiry or contact our Support Team ahead of purchase.

This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com

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.

Supplementary info

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

Activity summary

The protein p53 also known as TP53 or tumor protein p53 has a molecular weight of approximately 53 kDa. It acts as a transcription factor and plays a major role in cell cycle regulation apoptosis and maintaining genomic stability. This protein mainly expresses in the nucleus of cells and acts as a critical regulator of cellular responses to stress signals including DNA damage. Scientists commonly use p53 antibodies in various assays like western blot and p53 immunofluorescence to detect and study its expression and functional status in cells.

Biological function summary

P53 functions to control cell division and apoptosis serving as a guardian of the genome by preventing mutation accumulation. It does not form part of a larger complex under normal conditions but interacts with various other molecules to execute its functions. p53 can activate or suppress the transcription of numerous genes involved in cell cycle arrest DNA repair and programmed cell death allowing it to halt the progression of damaged cells and trigger repair mechanisms or eliminate those that cannot be repaired.

Pathways

P53 acts within several key biological pathways such as the p53 signaling pathway and the intrinsic apoptotic pathway. Its activity involves interaction with proteins like MDM2 which regulates p53 through ubiquitin-mediated degradation and ATM kinase which phosphorylates p53 in response to DNA damage. These interactions ensure appropriate cellular responses during stress and are vital for maintaining homeostasis.

Associated diseases and disorders

P53 mutation or inactivation is often associated with the development of cancer given its role in controlling cell division and preventing tumor formation. Specifically its dysfunction has been linked to cancers such as breast cancer and lung cancer. Additionally p53 can interact with other mutant proteins like Ras compounding mutations that contribute to tumor progression and aggressive cancer phenotypes. Understanding these interactions and the status of p53 can be important in developing targeted cancer therapies.

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.

12 product images

  • ChIP - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976), expandable thumbnail

    ChIP - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976)

    Chromatin was prepared from HEK-293 (human epithelial cell line from embryonic kidney) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25 μg of chromatin, 2 μg of Anti-p53 antibody [DO-1] - ChIP Grade ab1101, and 10 ml of protein A sepharose beads,10 ml of protein G sepharose beads. No IgG was added to the beads control. The immunoprecipitated DNA was quantified by real time PCR. Primers were as follows:

    Bax-1, forward: GGGTTATCTCTTGGGCTCACAA.

    Bax-1, reverse: GAGCTCTCCCCAGCGCA.

    Bax-2, forward: TGG AGC TGC AGA GGA TGA TTG

    Bax-2, reverse: CCA GTT GAA GTT GCC GTC AGA

    PUMA, forward: ATG CCT GCC TCA CCT TCA TC

    PUMA, reverse: TCA CAC GTC GCT CTC TCT AAA CC

    p21-1, forward: GCT GTG GCT CTG ATT GGC TTT

    p21-1, reverse: ACA GGC AGC CCA AGG ACA AA

    p21-2, forward: CAT CCC CAC AGC AGA GGA GAA

    p21-2, reverse: ACC CAG GCT TGG AGC AGC TA

    p21-3, forward: GAG TCC TGT TTG CTT CTG GGC A

    p21-3, reverse: CTG CAT TGG GGC TGC CTA TGT A

    PCNA, forward: CCA CCA TAA AGC TGG GGC TT

    PCNA, reverse: TCT CCC CGC CTC TTT GAC TC

    This image was produced using the same antibody clone but in a different formulation; PBS and sodium azide (Anti-p53 antibody [DO-1] - ChIP Grade ab1101).

  • Western blot - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976), expandable thumbnail

    Western blot - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976)

    False colour image of Western blot: Anti-p53 antibody [DO-1] - ChIP Grade staining at 1/1000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-p53 antibody [DO-1] - ChIP Grade ab1101 was shown to bind specifically to p53. A band was observed at 49 kDa in wild-type A549 cell lysates with no signal observed at this size in tp53 knockout cell line Human TP53 (p53) knockout A549 cell line ab276092 (knockout cell lysate ab282999). To generate this image, wild-type and tp53 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.

    All lanes: Western blot - Anti-p53 antibody [DO-1] - ChIP Grade (Anti-p53 antibody [DO-1] - ChIP Grade ab1101) at 1/1000 dilution

    Lane 1: Wild-type A549 cell lysate at 20 µg

    Lane 2: TP53 knockout A549 cell lysate at 20 µg

    Lane 3: Wild-type Jurkat cell lysate at 20 µg

    Lane 4: TP53 knockout Jurkat cell lysate at 20 µg

    Lane 5: A431 cell lysate at 20 µg

    Lane 6: Saos-2 cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 43 kDa

    Observed band size: 49 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976)

    Anti-p53 antibody [DO-1] - ChIP Grade ab1101 staining p53 in wild-type Hap1 cells (top panel) and p53 knockout Hap1 cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-p53 antibody [DO-1] - ChIP Grade ab1101 at 1μg/ml concentration and Anti-beta Tubulin antibody - Loading Control ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a high-content analysis system (Perkin Elmer, Operetta CLS™).

    This data was developed using the same antibody clone in a different buffer formulation (Anti-p53 antibody [DO-1] - ChIP Grade ab1101).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976)

    IHC image of Anti-p53 antibody [DO-1] - ChIP Grade ab1101 staining p53 in human colon adenocarcinoma formalin-fixed paraffin-embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with Anti-p53 antibody [DO-1] - ChIP Grade ab1101, 1/500 dilution, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. High magnification of the tumor region - T (lower right panel) and adjacent normal crypts - N (lower left panel) are shown.
    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.
    This image was produced using the same antibody clone but in a different formulation; PBS and sodium azide (Anti-p53 antibody [DO-1] - ChIP Grade ab1101).

  • Western blot - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976), expandable thumbnail

    Western blot - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976)

    False colour image of Western blot: Anti-p53 antibody [DO-1] - ChIP Grade staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-p53 antibody [DO-1] - ChIP Grade ab1101 was shown to bind specifically to p53. A band was observed at 50 kDa in wild-type HAP1 cell lysate with no signal observed at this size in tp53 knockout cell line. To generate this image, wild-type and tp53 knockout HAP1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-p53 antibody [DO-1] - ChIP Grade ab1101).

    All lanes: Western blot - Anti-p53 antibody [DO-1] - ChIP Grade (Anti-p53 antibody [DO-1] - ChIP Grade ab1101) at 1/1000 dilution

    Lane 1: Saos-2 cell lysate at 20 µg

    Lane 2: A431 cell lysate

    Lane 3: Wild-type HAP1 cell lysate at 20 µg

    Lane 4: TP53 knockout HAP1 cell lysate at 20 µg

    Lane 5: MCF7 cell lysate at 20 µg

    Lane 6: HEK-293T cell lysate at 20 µg

    Secondary

    Lanes 1 - 6: Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution

    Lanes 1 - 6: Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 43 kDa

    Observed band size: 50 kDa

    False colour image of Western blot: Anti-p53 antibody [DO-1] - ChIP Grade staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-p53 antibody [DO-1] - ChIP Grade ab1101 was shown to bind specifically to p53. A band was observed at 50 kDa in wild-type HAP1 cell lysate with no signal observed at this size in tp53 knockout cell line. To generate this image, wild-type and tp53 knockout HAP1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.

  • Western blot - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976), expandable thumbnail

    Western blot - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-p53 antibody [DO-1] - ChIP Grade ab1101).

    False colour image of Western blot: Anti-p53 antibody [DO-1] - ChIP Grade staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-p53 antibody [DO-1] - ChIP Grade ab1101 was shown to bind specifically to p53. A band was observed at 40/50 kDa in treated wild-type A549 cell lysates with no signal observed at this size in tp53 knockout cell line Human TP53 (p53) knockout A549 cell line ab276092. To generate this image, wild-type and tp53 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.

    Lanes 1, 10, 11, 12, 2, 3, 4, 5, 6, 7, 8 and 9: Western blot - Anti-p53 antibody [DO-1] - ChIP Grade (Anti-p53 antibody [DO-1] - ChIP Grade ab1101) at 1/1000 dilution

    Lanes 1, 10, 11, 12, 2, 3, 4, 5, 6, 7, 8 and 9: Western blot - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976)

    Lane 1: MCF7 Treated Camptothecin (20 ug/ml, 0 h) cell lysate at 15 µg

    Lane 2: MCF7 Treated Camptothecin (20 ug/ml, 6 h) cell lysate at 15 µg

    Lane 3: MCF7 Treated Camptothecin (20 ug/ml, 24 h) cell lysate at 15 µg

    Lane 4: Wild-type A549 Treated Camptothecin (20 ug/ml, 0 h) cell lysate at 15 µg

    Lane 5: Wild-type A549 Treated Camptothecin (20 ug/ml, 6 h) cell lysate at 15 µg

    Lane 6: Wild-type A549 Treated Camptothecin (20 ug/ml, 24 h) cell lysate at 15 µg

    Lane 7: TP53 knockout A549 Treated Camptothecin (20 ug/ml, 0 h) cell lysate at 15 µg

    Lane 8: TP53 knockout A549 Treated Camptothecin (20 ug/ml, 6 h) cell lysate at 15 µg

    Lane 9: TP53 knockout A549 Treated Camptothecin (20 ug/ml, 24 h) cell lysate at 15 µg

    Lane 10: Saos-2 cell lysate at 15 µg

    Lane 11: SK-BR-3 cell lysate at 15 µg

    Lane 12: A431 cell lysate at 15 µg

    Secondary

    All lanes: Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) at 1/20000 dilution

    Performed under reducing conditions.

  • Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-p53 antibody [DO-1] - ChIP Grade ab1101).

    Anti-p53 antibody [DO-1] - ChIP Grade ab1101 staining wild-type p53 in MCF7 cells (a low expressing cell line). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-p53 antibody [DO-1] - ChIP Grade ab1101 at 0.2µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

    Also suitable in cells fixed with 4% paraformaldehyde (10 min).

    Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

  • Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-p53 antibody [DO-1] - ChIP Grade ab1101).

    Anti-p53 antibody [DO-1] - ChIP Grade ab1101 staining wild-type p53 in Hek293 cells (a high expressing cell line). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-p53 antibody [DO-1] - ChIP Grade ab1101 at 0.2µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

    Also suitable in cells fixed with 4% paraformaldehyde (10 min).

    Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

  • Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-p53 antibody [DO-1] - ChIP Grade ab1101).

    Anti-p53 antibody [DO-1] - ChIP Grade ab1101 staining mutant p53 in A431 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-p53 antibody [DO-1] - ChIP Grade ab1101 at 0.2µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

    Also suitable in cells fixed with 100% methanol (5 min).

    Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

  • Flow Cytometry (Intracellular) - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-p53 antibody [DO-1] - ChIP Grade ab1101).

    Flow cytometry overlay histogram showing mutant p53 in A-431 cells stained with Anti-p53 antibody [DO-1] - ChIP Grade ab1101 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-p53 antibody [DO-1] - ChIP Grade ab1101) (1x 106 cells in 100μl at 0.04μg/ml (1/25000)) for 30min at 22°C. The secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) was incubated at 1/4000 for 30min at 22°C. Isotype control antibody (black line) Mouse IgG2a, Kappa Monoclonal [MOPC-173] - Isotype Control - ChIP Grade (Mouse IgG2a, Kappa Monoclonal [MOPC-173] -Isotype Control - ChIP Grade ab18413) was used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. This antibody gave a positive signal in A-431 fixed with 80% methanol (5 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

  • Flow Cytometry (Intracellular) - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-p53 antibody [DO-1] - ChIP Grade ab1101).
    Flow cytometry overlay histogram showing mutant p53 in wild-type HAP1 (green line) and TP53 knockout HAP1 cells (red line) stained with Anti-p53 antibody [DO-1] - ChIP Grade ab1101. The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-p53 antibody [DO-1] - ChIP Grade ab1101) (1x 106 cells in 100μl at 0.04 μg/ml (1/25000)) for 30min at 22°C.The secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) was incubated at 1/4000 for 30min at 22°C. Isotype control antibody Mouse IgG2a, Kappa Monoclonal [MOPC-173] - Isotype Control - ChIP Grade (Mouse IgG2a, Kappa Monoclonal [MOPC-173] -Isotype Control - ChIP Grade ab18413) was used at the same concentration and conditions as the primary antibody (wild-type HAP1 - black line, TP53 knockout HAP1 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. This antibody gave a positive signal in HAP1 fixed with 80% methanol (5 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

  • Flow Cytometry (Intracellular) - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-p53 antibody [DO-1] - BSA and Azide free (ab237976)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-p53 antibody [DO-1] - ChIP Grade ab1101).

    Flow cytometry overlay histogram showing wild-type p53 in Hek-293 positive cells (left) and MCF7 negative cells (right) stained with Anti-p53 antibody [DO-1] - ChIP Grade ab1101 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-p53 antibody [DO-1] - ChIP Grade ab1101) (1x 106 cells in 100μl at 0.2μg/ml (1/5000)) for 30min at 22°C. The secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) was incubated at 1/4000 for 30min at 22°C. Isotype control antibody (black line) Mouse IgG2a, Kappa Monoclonal [MOPC-173] - Isotype Control - ChIP Grade (Mouse IgG2a, Kappa Monoclonal [MOPC-173] -Isotype Control - ChIP Grade ab18413) was used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. This antibody gave a positive signal in Hek-293 fixed with 80% methanol (5 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

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