MA5-14067

antibody from Invitrogen Antibodies

Targeting: TP53 LFS1, p53

Provider product page for MA5-14067

Western blot

Immunohistochemistry

Flow cytometry

Chromatin Immunoprecipitation

Other assay

Antibody data

Antibody Data
Antigen structure
References [135]
Comments [0]
Validations
Western blot [2]
Immunohistochemistry [4]
Flow cytometry [1]
Chromatin Immunoprecipitation [1]
Other assay [5]

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Product number: MA5-14067 - Provider product page
Provider: Invitrogen Antibodies
Product name: p53 Antibody Cocktail
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MA5-14067 targets p53 in IHC (P), IP, and WB applications and shows reactivity with Human samples.
Antibody clone number: DO-7, BP53-12
Concentration: 0.125 mg/mL

TP53 protein structure - MA5-14067 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of HEL 92.1.7 (Lane 1), MCF-7 (Lane 2), MDA-MB-231 (Lane 3), COS-7 (Lane 4), A431 (Lane 5), and SKOV3 (Lane 6). The blots were probed with Anti-p53 Mouse Monoclonal Antibody (Product # MA5-14067, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 62-6520, 1:4000 dilution). A 53 kDa band corresponding to p53 was observed across cell lines tested except SKOV3. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of p53 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR718517_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of p53 was performed by loading 30 µg of HeLa wild type (Lane 1), HeLa Wild Type treated with 5 µM Camptothecin for 20hrs (Lane 2), HeLa Cas9 (Lane 3), HeLa Cas9 treated with 5 µM Camptothecin for 20hrs (Lane4), HeLa p53 KO (Lane 5) and HeLa p53 KO treated with 5 µM Camptothecin for 20hrs (Lane 6) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-p53 Antibody Cocktail (Product # MA5-14067, 1 µg/mL dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:5000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to p53.

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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of p53 showing staining in the nucleus and weak staining in the cytoplasm of paraffin-embedded human breast carcinoma tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval 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 Mouse Monoclonal Antibody (Product # MA5-14067) diluted in 3% BSA-PBS at a dilution of 1:50 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of p53 Antibody (DO-7 + BP53-12) showing staining in the nucleus of paraffin-embedded mouse colon tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval 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 Mouse Monoclonal Antibody (DO-7 + BP53-12) (Product # MA5-14067) diluted in 3% BSA-PBS at a dilution of 1:100 for 1 hour at 37°C in a humidified chamber. Tissues were washed extensively in 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of p53 showing staining in the nucleus and weak staining in the cytoplasm of paraffin-embedded human colon carcinoma tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval 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 Mouse Monoclonal Antibody (Product # MA5-14067) diluted in 3% BSA-PBS at a dilution of 1:20 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of p53 using p53 Monoclonal Antibody (Product # MA5-14067) on Native Human SKBR3 Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 P53, BRCA1 and BRCA2 staining in breast tumors . Signals were photographed in breast tissues at x 200. (A) Positive nuclear staining of p53 protein; (B) No obvious nuclear staining was observed over the whole normal breast tissue; (C) Positive nuclear staining of BRCA1 protein; (D) Negative control of BRCA1 staining in a breast tumor. (E) Positive nuclear staining of BRCA2 protein; (F) Negative control of BRCA2 staining in a breast tumor.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 2 Increased expression of miR-34a-activating pathways in cortical tubers. (A) Relative protein expression of pathological markers in cortical tubers compared with autopsy-derived controls; (B) representative western blots; (C-F) in situ hybridisation with double-labelling: (C) P-S6 was expressed by giant cells (arrowheads) and co-localised with miR-34a in situ hybridisation signal (IHS) to various extent; (D) expression of p53 with nuclear localisation in giant cells co-localised with miR-34a IHS (arrowhead); (E) expression of c-Myc co-localised with miR-34a IHS (arrowhead); (F) expression of p-JNK in control white matter (arrowhead), in giant cells in tuberous sclerosis complex (TSC) white matter (arrow), as well as in astrocytes in TSC grey matter (arrowheads); blue +/- sign indicate miR-34a IHS presence; red +/- indicate maker expression presence; scale bar in each panel is as indicated at the closest panel to the left, WM, white matter, GM, grey matter

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 Characterizing senescent cells. a , Representative images of CDKN1A (p21) and CDKN2A (p16) in A375 cells. Senescence was induced using doxorubicin (40 nM) for 4 days. The outline of the nucleus is indicated by the white line. Scale bar is 10 um. b , Scatterplot of the p21 nuclear intensity vs. the p16 nuclear intensity of individual cells either mock-treated (dividing) or doxorubicin-treated (senescent) A375 cells. Red dots indicate cells that were treated with doxorubicin. Units of fluorescence intensity are arbitrary. c , Representative images of TP53 (p53) in A375 cells treated as described in a. d , Box and whisker plots of p21, p16, and p53 nuclear intensities in A375 cells treated as described in a . **** p-value < 0.0001. e , Bar graph of the percent of A375 cells positive for SA-beta-gal. **** p-value < 0.0001. f , Scatterplot of the DNA content vs. the p21 nuclear intensity of individual A375 cells treated as in a . Color scale indicates the p16 nuclear intensity. Units of fluorescence intensity are arbitrary. The G2 phase cells are gated in blue. Percent positive for p21 and p16 is indicated for both dividing and senescent cells. See also Fig. S3. Fig 3

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 10.7717/peerj.12510/fig-6 Figure 6 FBXO9 inhibition facilitated the ferroptosis of osteoclasts by blocking the ubiquitin mediated-proteasome degradation of p53. (A) The target of FBXO9 was predicted by ubibrowser. (B) The p53 mRNA expression in the FBXO9 knockdown and control cell was assessed by qPCR ( n = 3). (C) The protein level of p53 in the FBXO9 knockdown and control cell was assessed by western blot ( n = 3). (D) FBXO9 directly interacts with p53. The proteins from BMDM derived osteoclasts were IP with IgG or antibodies against FBXO9 and p53, following by western blot analysis ( n = 3). (E) The stability of p53 protein was regulated by FBXO9. BMDM derived osteoclasts treated with or without si-FBXO9 in the presence of cycloheximide (CHX, 25 ug/ml) for various times as indicated and cell lysates were then assessed by western blot ( n = 3). ** p < 0.01. (F) The cell lysates isolated from scramble and si-FBXO9 infected BMDM derived osteoclasts were immunoprecipitated with anti-p53 antibody, then analyzed by western blot using ubiquitin antibody ( n = 3).