Anti-ATAD3A antibody [EPR27484-26]

13 product images

• 

  Western blot - Anti-ATAD3A antibody [EPR27484-26] (ab317252)

  This antibody recognizes overexpressed human ATAD3A isoform 2 and isoform 1 (UniProt ID: Q9NVI7).

  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-ATAD3A antibody [EPR27484-26] (ab317252) at 1/1000 dilution

  Lane 1: 293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a Human ATAD3A isoform 2 expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

  Lane 3: 293T cells transfected with a Human ATAD3A isoform 1 expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 66 kDa, 76 kDa, 36 kDa

  Exposure time: 62s

• 

  Western blot - Anti-ATAD3A antibody [EPR27484-26] (ab317252)

  This antibody does not cross-react with human ATAD3B and ATAD3C.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-ATAD3A antibody [EPR27484-26] (ab317252) at 1/1000 dilution

  Lane 1: His-tagged human ATAD3A fragment at 40 ng

  Lane 2: His-tagged human ATAD3B fragment at 40 ng

  Lane 3: His-tagged human ATAD3C fragment at 40 ng

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 36 kDa

  Exposure time: 180s

• 

  Western blot - Anti-ATAD3A antibody [EPR27484-26] (ab317252)

  Lanes 2-7 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 and lane 1 is applied with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  Exposure time: Lane 1: 26 seconds, lanes 2-7: 180 seconds

  All lanes: Western blot - Anti-ATAD3A antibody [EPR27484-26] (ab317252) at 1/1000 dilution

  Lane 1: Human cerebellum tissue lysate at 20 µg

  Lane 2: Mouse heart tissue lysate at 20 µg

  Lane 3: Mouse brain tissue lysate at 20 µg

  Lane 4: Mouse pancreas tissue lysate at 20 µg

  Lane 5: Rat heart tissue lysate at 20 µg

  Lane 6: Rat brain tissue lysate at 20 µg

  Lane 7: Rat spleen tissue lysate at 20 µg

  Secondary

  Lane 1: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

  Lanes 2 - 7: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 66 kDa, 36 kDa

• 

  Western blot - Anti-ATAD3A antibody [EPR27484-26] (ab317252)

  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  In Western blot, Anti-TOMM20 antibody [EPR15581-54] - Mitochondrial Marker (Anti-TOMM20 antibody [EPR15581-54] - Mitochondrial Marker ab186735) staining at 1/2000 dilution.

  All lanes: Western blot - Anti-ATAD3A antibody [EPR27484-26] (ab317252) at 1/1000 dilution

  Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) non-mitochondrial fraction at 20 µg

  Lane 2: HeLa mitochondria fraction at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 66 kDa, 36 kDa, 16 kDa

  Exposure time: 180s

• 

  Western blot - Anti-ATAD3A antibody [EPR27484-26] (ab317252)

  The lane 8 was developed using a high sensitivity ECL substrate.

  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  Exposure time: Lanes 1-4: 26 seconds, lanes 5-8: 180 seconds

  All lanes: Western blot - Anti-ATAD3A antibody [EPR27484-26] (ab317252) at 1/1000 dilution

  Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

  Lane 2: HeLa transfected with siRNA specifically targeting ATAD3A whole cell lysate at 20 µg

  Lane 3: A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 4: U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

  Lane 5: MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate at 20 µg

  Lane 6: 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 7: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

  Lane 8: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 66 kDa, 36 kDa

• 

  Flow Cytometry (Intracellular) - Anti-ATAD3A antibody [EPR27484-26] (ab317252)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling ATAD3A with ab317252 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

  Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

• 

  Flow Cytometry (Intracellular) - Anti-ATAD3A antibody [EPR27484-26] (ab317252)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling ATAD3A with ab317252 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

  Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-ATAD3A antibody [EPR27484-26] (ab317252)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling ATAD3A with ab317252 at 1/50 (10.12 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

  Confocal image showing mitochondrial staining in NIH/3T3 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
  
  Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

  anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain at 1/1000 (1ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-ATAD3A antibody [EPR27484-26] (ab317252)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling ATAD3A with ab317252 at 1/50 (10.12 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

  Confocal image showing mitochondrial staining in HeLa cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
  
  Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

  anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain at 1/1000 (1ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATAD3A antibody [EPR27484-26] (ab317252)

  Immunohistochemical analysis of paraffin-embedded Human colorectal carcinoma tissue labeling ATAD3A with ab317252 at 1/1000 (0.506 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on human colorectal carcinoma.
  
  The section was incubated with ab317252 for 30 mins at room temperature.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument

  Counterstained with Hematoxylin.

  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATAD3A antibody [EPR27484-26] (ab317252)

  Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling ATAD3A with ab317252 at 1/1000 (0.506 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on human breast cancer (PMID:36627348).
  
  The section was incubated with ab317252 for 30 mins at room temperature.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument

  Counterstained with Hematoxylin.

  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATAD3A antibody [EPR27484-26] (ab317252)

  Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling ATAD3A with ab317252 at 1/1000 (0.506 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on human cerebrum (PMID:35236834).
  
  The section was incubated with ab317252 for 30 mins at room temperature.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument

  Counterstained with Hematoxylin.

  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• 

  Immunoprecipitation - Anti-ATAD3A antibody [EPR27484-26] (ab317252)

  ATAD3A was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab317252 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317252 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

  Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
  
  Lane 2: ab317252 IP in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
  
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab317252 in HeLa whole cell lysate

  All lanes: Immunoprecipitation - Anti-ATAD3A antibody [EPR27484-26] (ab317252) at 1/30 dilution

  All lanes: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Exposure time: 6s