- MA5-26830 - Invitrogen Antibodies TP53BP1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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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 Monoclonal Antibody (OTI2H6) (Product # MA5-26830, 1:2000 ) 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-53BP1 Monoclonal Antibody (OTI2H6) (Product # MA5-26830) and a ~270kDa band corresponding to TP53-binding protein 1 was observed across cell lines tested . Nuclear enriched extracts (30 µg lysate) of HeLa (Lane 1), A-549 (Lane 2), MDA-MB-231 (Lane 3), MCF7 (Lane 4), U-2 OS (Lane 5), K-562 (Lane 6), U-87 MG (Lane 7) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of TP53-binding protein 1 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with 53BP1 Monoclonal Antibody (OTI2H6) (Product # MA5-26830) at 1:200 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded human breast tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded human thyroid tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded carcinoma of human thyroid tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded human endometrium tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded adenocarcinoma of human endometrium tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded carcinoma of human bladder tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded human lymph node tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded human lymphoma tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on paraffin-embedded human tonsil tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on paraffin-embedded adenocarcinoma of human breast tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on paraffin-embedded human colon tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on paraffin-embedded adenocarcinoma of human colon tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on paraffin-embedded human lung tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on paraffin-embedded carcinoma of human lung tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on paraffin-embedded human ovary tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on paraffin-embedded adenocarcinoma of human ovary tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on paraffin-embedded human pancreas tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a TP53BP1 monoclonal antibody (Product # MA5-26830) at a dilution of 1:150.

MA5-26830

Antibody data