Anti-BCAT2 antibody [EPR27488-72] - BSA and Azide free

10 product images

• 

  Immunoprecipitation - Anti-BCAT2 antibody [EPR27488-72] - BSA and Azide free (ab307834)

  This data was produced using Anti-BCAT2 antibody [EPR27488-72] ab307833, the same clone but in a different formulation.
  BCAT2 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with Anti-BCAT2 antibody [EPR27488-72] ab307833 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-BCAT2 antibody [EPR27488-72] ab307833 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: NIH/3T3 whole cell lysate 10 ug
  
  Lane 2: Anti-BCAT2 antibody [EPR27488-72] ab307833 IP in NIH/3T3 whole cell lysate
  
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-BCAT2 antibody [EPR27488-72] ab307833 in NIH/3T3 whole cell lysate
  
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.
  
  Exposure time: 3 seconds.

• 

  Immunoprecipitation - Anti-BCAT2 antibody [EPR27488-72] - BSA and Azide free (ab307834)

  This data was produced using Anti-BCAT2 antibody [EPR27488-72] ab307833, the same clone but in a different formulation.
  BCAT2 was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate with Anti-BCAT2 antibody [EPR27488-72] ab307833 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-BCAT2 antibody [EPR27488-72] ab307833 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: MCF7 whole cell lysate 10 ug
  
  Lane 2: Anti-BCAT2 antibody [EPR27488-72] ab307833 IP in MCF7 whole cell lysate
  
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-BCAT2 antibody [EPR27488-72] ab307833 in MCF7 whole cell lysate
  
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.
  
  Exposure time: Lanes 1-3: 6 seconds (left), Lanes 1-3: 180 seconds (right).
  
  The IP experiment was performed by Anti-BCAT2 antibody [EPR27488-72] ab307833 using MCF cells. On the left the IP blot was probed with Anti-BCAT2 antibody [EPR27488-72] ab307833 and on the right the blot was probed by another anti-BCAT2 antibody (Anti-BCAT2 antibody ab95976)(1:1000 dilution).

• 

  Flow Cytometry (Intracellular) - Anti-BCAT2 antibody [EPR27488-72] - BSA and Azide free (ab307834)

  This data was produced using Anti-BCAT2 antibody [EPR27488-72] ab307833, the same clone but in a different formulation.
  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 mouse embryonic fibroblast cells labelling BCAT2 with Anti-BCAT2 antibody [EPR27488-72] ab307833 at 1/500 dilution (0.1ug) (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). A 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-BCAT2 antibody [EPR27488-72] - BSA and Azide free (ab307834)

  This data was produced using Anti-BCAT2 antibody [EPR27488-72] ab307833, the same clone but in a different formulation.
  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labelling BCAT2 with Anti-BCAT2 antibody [EPR27488-72] ab307833 at 1/500 dilution (1.1 ug/ml) 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 dilution (2 ug/ml) (Green).
  
  Confocal image showing mitochondrial staining in MCF7 cell line.
  
  Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). Alexa Fluor® 647 Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker ab197491 Anti-COX IV Mouse monoclonal antibody (Alexa Fluor® 647) - Mitochondrial Marker was used to counterstain tubulin at 1/200 dilution (2.5 ug/ml)(Red). 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 dilution (2 ug/ml).

• 

  Immunocytochemistry/ Immunofluorescence - Anti-BCAT2 antibody [EPR27488-72] - BSA and Azide free (ab307834)

  This data was produced using Anti-BCAT2 antibody [EPR27488-72] ab307833, the same clone but in a different formulation.
  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling BCAT2 with Anti-BCAT2 antibody [EPR27488-72] ab307833 at 1/500 dilution (1.1 ug/ml) 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 dilution (2 ug/ml) (Green).
  
  Confocal image showing cytoplasmic staining in NIH/3T3 cell line.
  
  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 dilution (2.5 ug/ml) (Red). 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 dilution (2 ug/ml).

• 

  Western blot - Anti-BCAT2 antibody [EPR27488-72] - BSA and Azide free (ab307834)

  This data was produced using Anti-BCAT2 antibody [EPR27488-72] ab307833, the same clone but in a different formulation.
  
  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  The expression of BCAT2 is upregulated in response to 3T3-L1 differentiation for 7 days treatment (PMID:27689828).
  
  In Western blot, anti- Vinculin antibody (Anti-Vinculin antibody [EPR8185] ab129002) loading control staining at 1/10000 dilution.
  
  Exposure time: 6 seconds.

  All lanes: Western blot - Anti-BCAT2 antibody [EPR27488-72] (Anti-BCAT2 antibody [EPR27488-72] ab307833) at 1/1000 dilution

  Lane 1: Untreated 3T3-L1 whole cell lysate at 20 µg

  Lane 2: 3T3-L1 differentiation for 7 days 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 1/20000 dilution

  Observed band size: 37 kDa

  Exposure time: 6s

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  The expression of BCAT2 is upregulated in response to 3T3-L1 differentiation for 7 days treatment (PMID:27689828).
  
  In Western blot, anti- Vinculin antibody (Anti-Vinculin antibody [EPR8185] ab129002) loading control staining at 1/10000 dilution.
  
  Exposure time: 6 seconds.

• 

  Western blot - Anti-BCAT2 antibody [EPR27488-72] - BSA and Azide free (ab307834)

  This data was produced using Anti-BCAT2 antibody [EPR27488-72] ab307833, the same clone but in a different formulation.
  
  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
  
  Exposure time: 15 seconds.

  All lanes: Western blot - Anti-BCAT2 antibody [EPR27488-72] (Anti-BCAT2 antibody [EPR27488-72] ab307833) at 1/1000 dilution

  Lane 1: Mouse stomach tissue lysate at 20 µg

  Lane 2: Human stomach tissue lysate at 20 µg

  Lane 3: Rat stomach tissue 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: 37 kDa

  Exposure time: 15s

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
  
  Exposure time: 15 seconds.

• 

  Western blot - Anti-BCAT2 antibody [EPR27488-72] - BSA and Azide free (ab307834)

  This data was produced using Anti-BCAT2 antibody [EPR27488-72] ab307833, the same clone but in a different formulation.
  
  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  Exposure time: Lane 1, 4-5: 15 seconds, Lane 2-3: 81 seconds.

  All lanes: Western blot - Anti-BCAT2 antibody [EPR27488-72] (Anti-BCAT2 antibody [EPR27488-72] ab307833) at 1/1000 dilution

  Lane 1: Human skeletal muscle tissue lysate at 20 µg

  Lane 2: Human liver tissue lysate at 20 µg

  Lane 3: Mouse liver tissue lysate at 20 µg

  Lane 4: Mouse heart tissue lysate at 20 µg

  Lane 5: Mouse skeletal muscle tissue 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: 37 kDa

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  Exposure time: Lane 1, 4-5: 15 seconds, Lane 2-3: 81 seconds.

• 

  Western blot - Anti-BCAT2 antibody [EPR27488-72] - BSA and Azide free (ab307834)

  This data was produced using Anti-BCAT2 antibody [EPR27488-72] ab307833, the same clone but in a different formulation.
  
  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  In Western blot, anti- Vinculin antibody (Anti-Vinculin antibody [EPR8185] ab129002) loading control staining at 1/10000 dilution.
  
  Exposure time: 15 seconds.

  All lanes: Western blot - Anti-BCAT2 antibody [EPR27488-72] (Anti-BCAT2 antibody [EPR27488-72] ab307833) at 1/1000 dilution

  Lane 1: MCF7 whole cell lysate at 20 µg

  Lane 2: Jurkat whole cell lysate at 20 µg

  Lane 3: Raji whole cell lysate at 20 µg

  Lane 4: HeLa whole cell lysate at 20 µg

  Lane 5: NIH/3T3 whole cell lysate at 20 µg

  Lane 6: RAW264.7 whole cell lysate at 20 µg

  Lane 7: PC-12 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: 37 kDa

  Exposure time: 15s

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  In Western blot, anti- Vinculin antibody (Anti-Vinculin antibody [EPR8185] ab129002) loading control staining at 1/10000 dilution.
  
  Exposure time: 15 seconds.

• 

  Western blot - Anti-BCAT2 antibody [EPR27488-72] - BSA and Azide free (ab307834)

  This data was produced using Anti-BCAT2 antibody [EPR27488-72] ab307833, the same clone but in a different formulation.
  
  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  This antibody does not cross-react with human BCAT1.
  
  In Western blot, anti-His antibody (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.
  
  In Western blot, anti-H3 antibody (Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade ab176842) loading control staining at 1/100000 dilution.
  
  Exposure time: 1 second.

  All lanes: Western blot - Anti-BCAT2 antibody [EPR27488-72] (Anti-BCAT2 antibody [EPR27488-72] ab307833) at 1/1000 dilution

  Lane 1: 293T (human embryonic kidney epithelial cell)cells transfected with an empty vector containing a his tag whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a human BCAT2 expression vector containing a his tag whole cell lysate at 20 µg

  Lane 3: 293T cells transfected with a human BCAT1 expression vector containing a 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 1/20000 dilution

  Predicted band size: 44 kDa

  Observed band size: 37 kDa

  Exposure time: 1s

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  This antibody does not cross-react with human BCAT1.
  
  In Western blot, anti-His antibody (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.
  
  In Western blot, anti-H3 antibody (Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade ab176842) loading control staining at 1/100000 dilution.
  
  Exposure time: 1 second.