ATPB Recombinant Rabbit Monoclonal Antibody [JM10-90]
cat.: ET1703-29
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Zebrafish
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP
Clonality: Monoclonal
Clone number: JM10-90
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: 57 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human ATPB aa 141-184 / 529.
Positive control: Hela cell lysate, HepG2 cell lysate, 293T cell lysate, RH-35 cell lysate, mouse brain tissue lysate, mouse liver tissue lysate, mouse kidney tissue lysate, rat kidney tissue lysate, zebrafish tissue lysates, 293T, A431, C6, human liver tissue, human kidney tissue, human uterus tissue, mouse liver tissue, mouse brain tissue, mouse heart tissue.
Subcellular location: Mitochondrion. Mitochondrion inner membrane. Peripheral membrane protein.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  IP
1:1,000-1:5,000
1:100-1:500
1:100-1:500
1:50-1:200
1:10-1:50
Uniprot #: SwissProt: P06576 Human | P56480 Mouse | P10719 Rat
Alternative names: ATP 5B ATP synthase H+ transporting mitochondrial F1 complex beta polypeptide ATP synthase subunit beta mitochondrial ATP synthase subunit beta, mitochondrial atp5b ATPB ATPB_HUMAN ATPMB ATPSB Epididymis secretory protein Li 271 HEL-S-271 Mitochondrial ATP synthase beta subunit Mitochondrial ATP Synthase Subunit Beta Mitochondrial ATP synthetase beta subunit
Images
ET1703-29_1.jpg Fig1: Western blot analysis of ATPB on different lysates with Rabbit anti-ATPB antibody (ET1703-29) at 1/1,000 dilution.

Lane 1: Hela cell lysate
Lane 2: HepG2 cell lysate
Lane 3: 293T cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 57 kDa
Observed band size: 57 kDa

Exposure time: 2 minutes;

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 (ET1703-29) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
ET1703-29_2.jpg Fig2: Western blot analysis of ATPB on different lysates with Rabbit anti-ATPB antibody (ET1703-29) at 1/1,000 dilution.

Lane 1: RH-35 cell lysate (20 µg/Lane)
Lane 2: Mouse brain tissue lysate (40 µg/Lane)
Lane 3: Mouse liver tissue lysate (40 µg/Lane)
Lane 4: Mouse kidney tissue lysate (40 µg/Lane)
Lane 5: Rat kidney tissue lysate (40 µg/Lane)

Predicted band size: 57 kDa
Observed band size: 50 kDa

Exposure time: 24 seconds;

4-20% 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 (ET1703-29) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
ET1703-29_3.jpg Fig3: Western blot analysis of ATPB on zebrafish tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1703-29, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.
ET1703-29_4.jpg Fig4: Immunocytochemistry analysis of 293T cells labeling ATPB with Rabbit anti-ATPB antibody (ET1703-29) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-ATPB antibody (ET1703-29) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1703-29_5.jpg Fig5: Immunocytochemistry analysis of A431 cells labeling ATPB with Rabbit anti-ATPB antibody (ET1703-29) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-ATPB antibody (ET1703-29) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1703-29_6.jpg Fig6: Immunocytochemistry analysis of C6 cells labeling ATPB with Rabbit anti-ATPB antibody (ET1703-29) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-ATPB antibody (ET1703-29) at 1/100 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 (HA601187, 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.
ET1703-29_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-ATPB antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-29, 1/50) for 30 minutes 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.
ET1703-29_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-ATPB antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-29, 1/50) for 30 minutes 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.
ET1703-29_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded human uterus tissue using anti-ATPB antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-29, 1/50) for 30 minutes 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.
ET1703-29_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-ATPB antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-29, 1/50) for 30 minutes 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.
ET1703-29_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-ATPB antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-29, 1/50) for 30 minutes 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.
ET1703-29_12.jpg Fig12: Immunohistochemical analysis of paraffin-embedded mouse heart tissue using anti-ATPB antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-29, 1/50) for 30 minutes 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.
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