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Product Info Summary
| SKU: | A00397 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-53BP1/TP53BP1 Antibody Picoband®
SKU/Catalog Number
A00397
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-53BP1/TP53BP1 Antibody Picoband® catalog # A00397. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.
Cite This Product
Anti-53BP1/TP53BP1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00397)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human 53BP1/TP53BP1 recombinant protein (Position: H1239-H1972).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00397 is reactive to TP53BP1 in Human
Observed Molecular Weight
450 kDa
Calculated molecular weight
213.6 kDa
Background of TP53BP1
TP53BP1(Tumor Protein p53-Binding Protein 1), also called 53BP1, is a protein that in humans is encoded by the TP53BP1 gene. Iwabuchi et al.(1998) mapped the TP53BP1 gene to 15q15-q21 by FISH. Iwabuchi et al.(1994) showed that TP53BP1 binds to the conformationally sensitive central domain of wildtype p53 but not to mutant p53 in vitro. Immunoblot analysis by Iwabuchi et al.(1998) showed that expression of TP53BP1 or TP53BP2 enhances the transactivation function of p53 and induces the expression of p21(CDKN1A). Wang et al.(2002) used small interfering RNA directed against TP53BP1 in mammalian cells to demonstrate that TP53BP1 is a key transducer of the DNA damage checkpoint signal. TP53BP1 was required for p53 accumulation, G2/M checkpoint arrest, and the intra-S-phase checkpoint in response to ionizing radiation.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A00397 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5 μg/ml, Human
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human U20S whole cell
IHC: human bladder epithelial carcinoma tissue, human metaplasia of squamous cells of the renal pelvis tissue, human ovarian cancer tissue, human lung cancer tissue, human breast cancer tissue, human placenta tissue, human placenta tissue, human rectal moderately differentiated adenocarcinoma tissue
ICC/IF: Hela cell
FCM: Hela cell
Validation Images & Assay Conditions
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Western blot analysis of 53BP1/TP53BP1 using anti-53BP1/TP53BP1 antibody (A00397).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human U20S whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-53BP1/TP53BP1 antigen affinity purified polyclonal antibody (Catalog # A00397) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for 53BP1/TP53BP1 at approximately 450 kDa. The expected band size for 53BP1/TP53BP1 is at 214 kDa.
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Western blot analysis of 53BP1/TP53BP1 using anti-53BP1/TP53BP1 antibody (A00397).
Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human U2OS (Cell nucleus) whole cell lysates,
Lane 2: human U2OS (Cell nucleus) whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-53BP1/TP53BP1 antigen affinity purified polyclonal antibody (Catalog # A00397) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for 53BP1/TP53BP1 at approximately 450 kDa. The expected band size for 53BP1/TP53BP1 is at 214 kDa.
Click image to see more details
IHC analysis of 53BP1/TP53BP1 using anti-53BP1/TP53BP1 antibody (A00397).
53BP1/TP53BP1 was detected in a paraffin-embedded section of human bladder epithelial carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-53BP1/TP53BP1 Antibody (A00397) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
IHC analysis of 53BP1/TP53BP1 using anti-53BP1/TP53BP1 antibody (A00397).
53BP1/TP53BP1 was detected in a paraffin-embedded section of human metaplasia of squamous cells of the renal pelvis tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-53BP1/TP53BP1 Antibody (A00397) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
IHC analysis of 53BP1/TP53BP1 using anti-53BP1/TP53BP1 antibody (A00397).
53BP1/TP53BP1 was detected in a paraffin-embedded section of human ovarian cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-53BP1/TP53BP1 Antibody (A00397) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
IHC analysis of 53BP1/TP53BP1 using anti-53BP1/TP53BP1 antibody (A00397).
53BP1/TP53BP1 was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-53BP1/TP53BP1 Antibody (A00397) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
IHC analysis of 53BP1/TP53BP1 using anti-53BP1/TP53BP1 antibody (A00397).
53BP1/TP53BP1 was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-53BP1/TP53BP1 Antibody (A00397) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
IHC analysis of 53BP1/TP53BP1 using anti-53BP1/TP53BP1 antibody (A00397).
53BP1/TP53BP1 was detected in a paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-53BP1/TP53BP1 Antibody (A00397) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
IHC analysis of 53BP1/TP53BP1 using anti-53BP1/TP53BP1 antibody (A00397).
53BP1/TP53BP1 was detected in a paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-53BP1/TP53BP1 Antibody (A00397) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
IHC analysis of 53BP1/TP53BP1 using anti-53BP1/TP53BP1 antibody (A00397).
53BP1/TP53BP1 was detected in a paraffin-embedded section of human rectal moderately differentiated adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-53BP1/TP53BP1 Antibody (A00397) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
IF analysis of 53BP1/TP53BP1 using anti-53BP1/TP53BP1 antibody (A00397).
53BP1/TP53BP1 was detected in an immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-53BP1/TP53BP1 Antibody (A00397) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Flow Cytometry analysis of Hela cells using anti-53BP1/TP53BP1 antibody (A00397).
Overlay histogram showing Hela cells stained with A00397 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-53BP1/TP53BP1 Antibody (A00397, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-53BP1/TP53BP1 Antibody Picoband® (A00397)
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