MA5-15244

antibody from Invitrogen Antibodies

Targeting: TP53 LFS1, p53

Antibody data

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Validation data

Product number: MA5-15244 - Provider product page
Provider: Invitrogen Antibodies
Product name: p53 Monoclonal Antibody (PAb 240)
Antibody type: Monoclonal
Antigen: Other
Description: MA5-15244 targets p53 in FACS, IHC (P), IP, and WB applications and shows reactivity with Human samples.
Antibody clone number: PAb 240
Concentration: 0.2 mg/mL

TP53 protein structure - MA5-15244 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot of p53 using p53 Monoclonal Antibody (Product # MA5-15244) on MDA231 Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of COS-7 (Lane 1), A431 (Lane 2), and SK-OV-3 (Lane 3). The blots were probed with Anti-p53 Mouse Monoclonal Antibody (Product # MA5-15244, 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 SK-OV-3. 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)
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Experimental details: Knockout of P53 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR718498_LV and 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 CAS9 (Lane 2) and HeLa P53 KO (Lane 3) whole cell extracts. The samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 Monoclonal Antibody (PAb 240) (Product # MA5-15244) using 1 µg/mL dilution and Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to P53. Uncharacterized bands were observed at ~30 kDa and 60 kDa in all the samples and ~80 kDa in HeLa CAS9 sample.