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Western blot
ImmunohistochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
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- Product number
- PA5-22758 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- 53BP1 Polyclonal Antibody, DyLight™ 650
- Antibody type
- Polyclonal
- Antigen
- Other
- Description
- Human samples have been tested in Western Blot and ICC/IF, and mouse samples have been tested in ICC/IF only. Suggested positive control: U205, 293T, and MO59K cell lysates and MEF lysates .
- Reactivity
- Human, Mouse, Rat, Bovine, Canine
- Host
- Rabbit
- Conjugate
- Red dye
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.8 mg/mL
- Storage
- 4° C, store in dark
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-53BP1 Polyclonal Antibody, DyLight 650 (Product # PA5-22758) and a ~270 kDa band corresponding to TP53-binding protein 1 was observed across cell lines tested. Nuclear enriched extracts (50 µg lysate) of HeLa (Lane 1), MCF7 (Lane 2), U-2 OS (Lane 3), K-562 (Lane 4), U-87 MG (Lane 5) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB24001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescentdetection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).A streak like pattern was observed in the positive cell lines as expected of 53BP1 target.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of TP53-binding protein 1 was achieved by transfecting HeLa with TP53-binding protein 1 specific siRNAs (Silencer® select Product # s14315, s14314). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the TP53-binding protein 1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with 53BP1 Polyclonal Antibody, DyLight 650 (Product # PA5-22758, 1:1000 dilution ) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to TP53-binding protein 1.A streak like pattern was observed in the positive cell lines as expected of 53BP1 target.
