MA5-12554

antibody from Invitrogen Antibodies

Targeting: TP53 LFS1, p53

Provider product page for MA5-12554

Western blot

Antibody data

Antibody Data
Antigen structure
References [138]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [4]
Immunohistochemistry [3]
Flow cytometry [3]
Other assay [2]

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Product number: MA5-12554 - Provider product page
Provider: Invitrogen Antibodies
Product name: p53 Monoclonal Antibody (DO-7), Biotin
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MA5-12554 targets p53 in ICC/IF, FACS, IHC (P), IP, and WB applications and shows reactivity with Bovine, Human, and Non-human primate samples.
Conjugate: Biotin
Antibody clone number: DO-7
Concentration: 0.2 mg/mL

TP53 protein structure - MA5-12554 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of p53 (green) showing staining in the nucleus of A431 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a p53 monoclonal antibody (Product # MA5-12557) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Activation of ATM and p53: ATM kinase is auto-phosphorylated in response to DNA damage and is involved in DNA damage induced activation of p53 and cell cycle arrest. This is a multiplex kit, enabling the user to simultaneously measure both phospho-ATM and p53. A549 cells were treated with vehicle (DMSO) or 50 µM etoposide for 24 hrs. After staining cells with Cellomics phospho-ATM/p53 duplex activation kit, images were acquired and analyzed. Z-factor for phospho-ATM = 0.46 ± 0.11 and %COV = 11 ± 3. Z-factor for p53 = 0.44 ± 0.03 and %COV = 13 ± 1. EC50 of etoposide treatment for ATM activation is 19 µM and EC50 for p53 activation is 36 µM. Etoposide is a topoisomerase II inhibitor resulting in DNA strand break.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of p53 (green) showing staining in the nucleus of HT29 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a p53 monoclonal antibody (Product # MA5-12557) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of p53 (green) in HeLa cells either left untreated (left panel) or treated with 5uM Camptothecin (right panel) for 20 hours. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a p53 monoclonal antibody (Product # MA5-12557) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or ToxInsight Instrument at 20X magnification.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Formalin-fixed, paraffin-embedded human colon carcinoma stained with p53 antibody using peroxidase-conjugate and DAB chromogen. Note nuclear staining of tumor cells.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of p53 showing positive staining in the nucleus and cytoplasm of paraffin-treated Human breast carcinoma (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a p53 monoclonal antibody (Product # MA5-12557) diluted by 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of p53 showing positive staining in the nucleus and cytoplasm of paraffin-treated Human colon carcinoma (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a p53 monoclonal antibody (Product # MA5-12557) diluted by 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of p53 in A431 cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde, washed with PBS, and incubated with p53 monoclonal antibody (Product # MA5-12557) at a dilution of 0.5 µg/test for 60 min at room temperature. Cells were then blocked in a solution of 2% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody, and re-suspended in PBS for FACS analysis.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of p53 in BAEC cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde, washed with PBS, and incubated with p53 monoclonal antibody (Product # MA5-12557) at a dilution of 0.5 µg/test for 60 min at room temperature. Cells were then blocked in a solution of 2% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody, and re-suspended in PBS for FACS analysis.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of p53 in CEM cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde, washed with PBS, and incubated with p53 monoclonal antibody (Product # MA5-12557) at a dilution of 1 µg/test for 60 min at room temperature. Cells were then blocked in a solution of 2% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody, and re-suspended in PBS for FACS analysis.
Conjugate: Biotin