Name: Anti-p53 antibody [PAb 240]
SKU: ab26
Rating: 4 (55 reviews)

Anti-p53 antibody [PAb 240] (ab26) is a mouse monoclonal antibody detecting p53 in Western Blot, IP, ICC/IF. Suitable for Human, Mouse.

- KO validated for confirmed specificity

- Over 400 publications

- Trusted since 1998

Images

Western blot - Anti-p53 antibody [PAb 240] (AB26), expandable thumbnail

Publications

Key facts

Isotype: IgG1
Host species: Mouse
Storage buffer: pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Form: Liquid
Clonality: Monoclonal

Immunogen

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

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF
Human	Tested	Tested	Tested
Mouse	Tested	Tested	Tested
Rat	Predicted	Predicted	Predicted
Chinese hamster	Predicted	Predicted	Predicted
Cow	Predicted	Predicted	Predicted
Dog	Predicted	Predicted	Predicted
Monkey	Predicted	Predicted	Predicted
Syrian hamster	Predicted	Predicted	Predicted

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse	Dilution info -	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Rat, Cow, Dog, Monkey, Chinese hamster, Syrian hamster	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1.00000-5.00000 µg/mL	Notes Please note that expression of target protein may be very low without stimulation/treatment (e.g. DNA damaging agent). We recommend using 3% milk as the blocking agent for Western blot.
Species Mouse	Dilution info 1.00000-5.00000 µg/mL	Notes Please note that expression of target protein may be very low without stimulation/treatment (e.g. DNA damaging agent). We recommend using 3% milk as the blocking agent for Western blot.

Predicted
Predicted

Species	Dilution info	Notes
Species Rat, Cow, Dog, Monkey, Chinese hamster, Syrian hamster	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 0.50000-1.00000 µg/mL	Notes -
Species Mouse	Dilution info 0.50000-1.00000 µg/mL	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Rat, Cow, Dog, Monkey, Chinese hamster, Syrian hamster	Dilution info -	Notes -

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Target data

See full target information Tp53

Function

Multifunctional transcription factor that induces cell cycle arrest, DNA repair or apoptosis upon binding to its target DNA sequence (PubMed:19556538, PubMed:20673990, PubMed:22726440). Acts as a tumor suppressor in many tumor types; induces growth arrest or apoptosis depending on the physiological circumstances and cell type. Negatively regulates cell division by controlling expression of a set of genes required for this process. 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. Its pro-apoptotic activity is activated via its interaction with PPP1R13B/ASPP1 or TP53BP2/ASPP2 (By similarity). However, this activity is inhibited when the interaction with PPP1R13B/ASPP1 or TP53BP2/ASPP2 is displaced by PPP1R13L/iASPP (By similarity). In cooperation with mitochondrial PPIF is involved in activating oxidative stress-induced necrosis; the function is largely independent of transcription. Prevents CDK7 kinase activity when associated to CAK complex in response to DNA damage, thus stopping cell cycle progression (By similarity). 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, but seems to have to effect on cell-cycle regulation. Regulates the circadian clock by repressing CLOCK-BMAL1-mediated transcriptional activation of PER2 (PubMed:24051492).

Alternative names

P53, Trp53, Tp53, Cellular tumor antigen p53, Tumor suppressor p53

Key facts

Isotype: IgG1
Host species: Mouse
Storage buffer: pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: PAb 240
Purification technique: Affinity purification
Epitope: The epitope has been mapped to between amino acids 212 and 217 on human p53 (PMID: 1376364).
Light chain type: kappa
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

ab26 has been knockout validated in Western blot. The expected band was seen in wild type HCT116 cells treated with the DNA damaging agent irinotecan and no band was seen in TP53 knockout HCT116 cells.

Product Specifications
Anti-p53 antibody [PAb 240] (ab26) is a mouse monoclonal antibody and is validated for use in ICC/IF, IP, WB in human, mouse samples.
Anti-p53 antibody [PAb 240] (ab26) specifically detects p53 (UniProt ID: P04637; Molecular weight: 44kDa) and is sold in 100 µg selling sizes.

Quality and Validation
Abcam's high quality validation processes ensure Anti-p53 antibody [PAb 240] (ab26) has high sensitivity and specificity.
The specificity of Anti-p53 antibody [PAb 240] (ab26) has been confirmed by testing in knockout samples.
Anti-p53 antibody [PAb 240] (ab26) has been cited over 403 times in peer reviewed journals and is trusted by the scientific community.
Anti-p53 antibody [PAb 240] (ab26) has 54 independent reviews from customers.

Related Products
Conjugation-ready, carrier free format available for antibody clone PAb 240 - ab185238.

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

Product promise

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

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8 product images

Western blot - Anti-p53 antibody [PAb 240] (ab26)
Lanes 1-6: Merged (red and green) signal.

ab26 was shown to specifically react with p53 in wild type HCT116 cells treated with irinotecan. No band was observed in p53 knockout HCT116 cells. Wild-type and p53 knockout samples, positive and negative controls were subjected to SDS-PAGE. ab26 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (loading control to GAPDH) were diluted 5 μg/mL and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging.
Wild-type and p53 knockout HCT116 cell lysates were kindly provided by a collaborator.
All lanes: Western blot - Anti-p53 antibody [PAb 240] (ab26) at 5 µg/mL
Lane 1: Wild-type HCT116 cell lysate at 30 µg
Lane 2: Wild-type HCT116 + irinotecan (10 µM, 24 hours) cell lysate at 30 µg
Lane 3: p53 knockout HCT116 cell lysate at 30 µg
Lane 4: p53 knockout HCT116 + irinotecan (10 µM, 24 hours) cell lysate at 30 µg
Lane 5: A431 cell lysate (positive control) at 20 µg
Lane 6: Saos-2 cell lysate (negative control) at 20 µg
Performed under reducing conditions.
Predicted band size: 43 kDa
Observed band size: 53 kDa
This image is courtesy of a customer review submitted by Dr Cherie Blenkiron
Western blot - Anti-p53 antibody [PAb 240] (ab26)
All lanes: Western blot - Anti-p53 antibody [PAb 240] (ab26) at 1/2000 dilution
Lane 1: Human breast cancer cell-line, MCF7 cells (p53 WT), whole cell lysate at 20 µg
Lane 2: Human breast cancer cell-line, MDA231 cells (p53 Mutant), whole cell lysate at 20 µg
Secondary
All lanes: HRP conjugated donkey anti-mouse antibody at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 43 kDa
Observed band size: 53 kDa, 72 kDa
Exposure time: 10s
Immunoprecipitation - Anti-p53 antibody [PAb 240] (ab26)
p53 was immunoprecipitated from 7x10⁶ HCT116 (human colon carcinoma cell line) cells with ab26 at 1/150 dilution. Western blot was performed from the immunoprecipitate using anti-p53 antibody. Donkey Anti-Mouse IgG H&L (Alexa Fluor® 750) preadsorbed (Donkey Anti-Mouse IgG H&L (Alexa Fluor® 750) preadsorbed ab175739) was used as secondary antibody at 1/5000 dilution.

Lane 1: HCT116 whole cell lysate 10 μg (Input).
Lane 2: Anti-Adipose Triglyceride Lipase antibody [EPR19650] ab207799 IP in etoposide treated HCT116 whole cell lysate.
Lane 3: Anti-Adipose Triglyceride Lipase antibody [EPR19650] ab207799 IP in etoposide treated HCT116 p53-/- whole cell lysate (negative control).
All lanes: Anti p53 antibody
Secondary
All lanes: Immunoprecipitation - Donkey Anti-Mouse IgG H&L (Alexa Fluor® 750) preadsorbed (Donkey Anti-Mouse IgG H&L (Alexa Fluor® 750) preadsorbed ab175739) at 1/5000 dilution
Predicted band size: 43 kDa
Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [PAb 240] (ab26)
ab26 stained in A431 cells. Cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with ab26 at 1μg/ml and Anti-beta Tubulin antibody - Loading Control ab6046 (Rabbit polyclonal to beta tubulin) at 1ug/ml overnight at +4°C. The secondary antibodies were Donkey Anti-Sheep IgG H&L (Alexa Fluor® 488) ab150177 (colored green) used at 1 ug/ml and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed ab150087 (pseudo-colored red) used at 2ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43μM for 1hour at room temperature.
Western blot - Anti-p53 antibody [PAb 240] (ab26)
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab26 overnight at 4°C. Antibody binding was detected using an anti-mouse HRP secondary antibody (Goat Anti-Mouse IgG H&L (HRP) preadsorbed ab97040), and visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406.
All lanes: Western blot - Anti-p53 antibody [PAb 240] (ab26) at 1 µg/mL
All lanes: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (Goat Anti-Mouse IgG H&L (HRP) preadsorbed ab97040) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 43 kDa
Observed band size: 53 kDa
Exposure time: 8min
Western blot - Anti-p53 antibody [PAb 240] (ab26)
All lanes: Western blot - Anti-p53 antibody [PAb 240] (ab26) at 5 µg/mL
Lane 1: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg
Lane 2: Hela Whole Cell Lysate - Bleomycin Treated (40U/ml) at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (Goat Anti-Mouse IgG H&L (HRP) preadsorbed ab97040) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 43 kDa
Exposure time: 4min
Western blot - Anti-p53 antibody [PAb 240] (ab26)
Lanes 1-2: 1% BSA blocking buffer
Lanes 3-4: 3% Milk blocking buffer
We recommend using 3% milk as the blocking agent for Western blot.
Lanes 1 - 2: Western blot - Anti-p53 antibody [PAb 240] (ab26) at 1 µg/mL
Lanes 3 - 4: Western blot - Anti-p53 antibody [PAb 240] (ab26) at 5 µg/mL
All lanes: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg
Secondary
All lanes: Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 43 kDa
Exposure time: 4min
Western blot - Anti-p53 antibody [PAb 240] (ab26)
Primary: All Lanes: Anti-p53 antibody (ab26) at 5 μg/mL. Lane 1: MW marker. Lane 2: NIH/3T3 cells treated with vehicle for 24 hours. Lane 3: NIH/3T3 cells treated with 1 μM doxorubicin for 24 hours Secondary: All Lanes: HRP-conjugated VeriBlot anti-Mouse IgG (Anti-mouse IgG for IP (HRP) ab131368) 1:1000. Lysates at 20 μg/lane. Performed under denaturing conditions. Developed using ECL technique. Blocking buffer: 5% milk in PBS.
All lanes: Western blot - Anti-p53 antibody [PAb 240] (ab26)
Predicted band size: 43 kDa