53BP1 Recombinant Rabbit Monoclonal Antibody [JA66-11]
cat.: ET1704-05
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: JA66-11
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 214 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human 53BP1 aa 1-50 / 1972.
Positive control: HeLa cell lysate, HT-29 cell lysate, human brain tissue lysate, mouse brain tissue lysate, rat heart tissue lysate, HeLa, human brain tissue, mouse spleen tissue, mouse testis tissue, rat testis tissue, human spleen tissue, human liver tissue, human cervix tissue.
Subcellular location: Nucleus. Chromosome.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:2,000
1:50
1:200
1:200
1:1,000
Uniprot #: SwissProt: Q12888 Human | P70399 Mouse
Entrez Gene: 296099 Rat
Alternative names: 53 BP1 53BP1 FLJ41424 MGC138366 p202 p53 binding protein 1 p53 BP1 p53-binding protein 1 p53BP1 TP53 BP1 TP53B_HUMAN Tp53bp1 TRP53 BP1 Tumor protein 53 binding protein 1 Tumor protein p53 binding protein 1 Tumor suppressor p53 binding protein 1 Tumor suppressor p53-binding protein 1
Images
ET1704-05_1.jpg Fig1: Western blot analysis of 53BP1 on different lysates with Rabbit anti-53BP1 antibody (ET1704-05) at 1/2,000 dilution.

Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: HT-29 cell lysate (15 µg/Lane)
Lane 3: Human brain tissue lysate (20 µg/Lane)
Lane 4: Mouse brain tissue lysate (20 µg/Lane)
Lane 5: Rat heart tissue lysate (20 µg/Lane)

Predicted band size: 214 kDa
Observed band size: 450 kDa

Exposure time: 43 seconds; ECL: K1801;

3-8% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1704-05) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1704-05_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling 53BP1 with Rabbit anti-53BP1 antibody (ET1704-05) at 1/50 dilution.

Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-53BP1 antibody (ET1704-05) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1704-05_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-05) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1704-05_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-05) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1704-05_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-05) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1704-05_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-05) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1704-05_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-05) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1704-05_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-05) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1704-05_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded human cervix tissue with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-05) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1704-05_10.jpg Fig10: Immunofluorescence analysis of paraffin-embedded human brain tissue labeling 53BP1 with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1704-05, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
ET1704-05_11.jpg Fig11: Immunofluorescence analysis of paraffin-embedded mouse testis tissue labeling 53BP1 with Rabbit anti-53BP1 antibody (ET1704-05) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1704-05, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
ET1704-05_12.jpg Fig12: Flow cytometric analysis of HeLa cells labeling 53BP1.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1704-05, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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