Anti-AKR1C3 antibody [EPR28747-41]

13 product images

• 

  Western blot - Anti-AKR1C3 antibody [EPR28747-41] (ab316864)

  AKR1C3 Western blot staining using rabbit Anti-AKR1C3 antibody

  Low expression:HCT 116 (PMID: 21215737, PMID: 35388063)

  In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.

  All lanes: Western blot - Anti-AKR1C3 antibody [EPR28747-41] (ab316864) at 1/1000 dilution

  Lane 1: TF-1 (human Erythroleukemia erythroblast) whole cell lysate at 20 µg with 5% NFDM/TBST

  Lane 2: HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

  Lane 3: A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

  Lane 4: HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

  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, 124 kDa

  Exposure time: 10s

• 

  Dot Blot - Anti-AKR1C3 antibody [EPR28747-41] (ab316864)

  AKR1C3 Dot Blot staining using rabbit Anti-AKR1C3 antibody

  Dot blot analysis of AKR1C3 using ab316864 (Lane 1-4) at 1/1000 followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 Dilution] dilution and Dot blot analysis of AKR1C3 using Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204 (lane 5-8) at 1/1000 followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution.

  Lane 1&5: His tagged AKR1C1 recombinant protein
  

  Lane 2&6: His tagged AKR1C2 recombinant protein
  

  Lane 3&7: AKR1C3 recombinant protein without tag
  

  Lane 4&8: His tagged AKR1C4 recombinant protein
  

  Lanes 1 - 4: Dot Blot - Anti-AKR1C3 antibody [EPR28747-41] (ab316864) at 1/1000 dilution

  Lanes 5 - 8: Dot Blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) at 1/1000 dilution

  Lanes 1 and 5: His tagged AKR1C1 recombinant protein

  Lanes 2 and 6: His tagged AKR1C2 recombinant protein

  Lanes 3 and 7: AKR1C3 recombinant protein without tag

  Lanes 4 and 8: His tagged AKR1C4 recombinant protein

  Secondary

  All lanes: Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051)

  Exposure time: 180s

• 

  Western blot - Anti-AKR1C3 antibody [EPR28747-41] (ab316864)

  Low expression: muscle (PMID: 18635746)

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

  All lanes: Western blot - Anti-AKR1C3 antibody [EPR28747-41] (ab316864) at 1/1000 dilution

  Lane 1: Mouse colon tissue lysate at 20 µg with 5% NFDM/TBST

  Lane 2: Mouse kidney tissue lysate at 20 µg with 5% NFDM/TBST

  Lane 3: Mouse liver tissue lysate at 20 µg with 5% NFDM/TBST

  Lane 4: Mouse skeletal muscle tissue lysate at 20 µg with 5% NFDM/TBST

  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: 10s

• 

  Western blot - Anti-AKR1C3 antibody [EPR28747-41] (ab316864)

  Low expression: muscle (PMID: 18635746)

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

  All lanes: Western blot - Anti-AKR1C3 antibody [EPR28747-41] (ab316864) at 1/1000 dilution

  Lane 1: Human liver tissue lysate at 20 µg with 5% NFDM/TBST

  Lane 2: Human kidney tissue lysate at 20 µg with 5% NFDM/TBST

  Lane 3: Human testis tissue lysate at 20 µg with 5% NFDM/TBST

  Lane 4: Human skeletal muscle tissue lysate at 20 µg with 5% NFDM/TBST

  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: 8s

• 

  Immunoprecipitation - Anti-AKR1C3 antibody [EPR28747-41] (ab316864)

  AKR1C3 was immunoprecipitated from 0.35 mg A549 (human lung carcinoma epithelial cell) whole cell lysate with ab316864 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316864 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: A549 (human lung carcinoma epithelial cell) whole cell lysate
  
  Lane 2: ab316864 IP in A549 (human lung carcinoma epithelial cell) whole cell lysate
  
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab316864 in A549 whole cell lysate

  All lanes: Immunoprecipitation - Anti-AKR1C3 antibody [EPR28747-41] (ab316864) at 1/30 dilution

  All lanes: A549 (human lung carcinoma epithelial cell) whole cell lysate with 5% NFDM/TBST

  Secondary

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

  Exposure time: 6s

• 

  Flow Cytometry (Intracellular) - Anti-AKR1C3 antibody [EPR28747-41] (ab316864)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HCT 116 (human colorectal carcinoma epithelial cell, Left) / TF-1 (human erythroleukemia erythroblast, Right) cells labelling AKR1C3 with ab316864 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.

  Low expression: HCT 116.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-AKR1C3 antibody [EPR28747-41] (ab316864)

  Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized TF-1 (human erythroleukemia erythroblast) cells labelling AKR1C3 with ab316864 at 1/50 (10.0 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 nuclear and cytoplasmic staining in TF-1 cell line.
  
  Low expression: HCT 116 (PMID: 21215737).
  
  Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

  Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/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-AKR1C3 antibody [EPR28747-41] (ab316864)

  Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling AKR1C3 with ab316864 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Negative control: No staining on mouse cardiac muscle.
  
  The section was incubated with ab316864 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-AKR1C3 antibody [EPR28747-41] (ab316864)

  Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling AKR1C3 with ab316864 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Negative control: No staining on human cardiac muscle (PMID: 33352741).
  
  The section was incubated with ab316864 for 30 mins at room temperature.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument
  
  Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

  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-AKR1C3 antibody [EPR28747-41] (ab316864)

  Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling AKR1C3 with ab316864 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on mouse colon.
  
  The section was incubated with ab316864 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-AKR1C3 antibody [EPR28747-41] (ab316864)

  Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling AKR1C3 with ab316864 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on mouse liver.
  
  The section was incubated with ab316864 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-AKR1C3 antibody [EPR28747-41] (ab316864)

  Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling AKR1C3 with ab316864 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on human hepatocellular carcinoma.
  
  The section was incubated with ab316864 for 30 mins at room temperature.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument
  
  Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

  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-AKR1C3 antibody [EPR28747-41] (ab316864)

  Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling AKR1C3 with ab316864 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on human colon (PMID: 33352741).
  
  The section was incubated with ab316864 for 30 mins at room temperature.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument
  
  Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

  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