Anti-alpha 1 Fetoprotein antibody [RM1011]

9 product images

• 

  Immunocytochemistry - Anti-alpha 1 Fetoprotein antibody [RM1011] (ab284388)

  This data was developed using ab284388, the same antibody clone in a different buffer formulation.
  

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 cells labelling alpha 1 Fetoprotein with ab284388 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 (2μg/mL) dilution (Green). 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 2.5μg/mL dilution (Red). The Nuclear counterstain was DAPI (Blue).

  Confocal image showing cytoplasmic staining in HepG2 cells is observed.

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 (2μg/mL) dilution.

• 

  Immunocytochemistry - Anti-alpha 1 Fetoprotein antibody [RM1011] (ab284388)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Hepa1-6 cells labelling alpha 1 Fetoprotein with ab284388 at 1/50 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 2μg/mL dilution (Green). 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 (2.5μg/mL) dilution (Red). The Nuclear counterstain was DAPI (Blue).
  

  Confocal image showing increased cytoplasmic staining in Hepa1-6 cells treated with Brefeldin A (300 ng/ml) for 24 h is observed.

  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 (2μg/mL) dilution.

• 

  Flow Cytometry - Anti-alpha 1 Fetoprotein antibody [RM1011] (ab284388)

  Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized Hepa1-6 (Mouse hepatoma epithelial cell) treated with 300ng/ml BFA for 24h (Right) / Untreated control (Left) cells labelling alpha 1 Fetoprotein with ab284388 at 1/500 dilution. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Fetoprotein antibody [RM1011] (ab284388)

  Immunohistochemical analysis of paraffin-embedded Human hepatic carcinoma tissue labelling alpha 1 Fetoprotein with ab284388 at 1/2000 (0.231 μg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human hepatic carcinoma. The section was incubated with ab284388 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Fetoprotein antibody [RM1011] (ab284388)

  Negative control: No staining on human liver.
  

  Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling alpha 1 Fetoprotein with ab284388 at 1/2000 (0.231 μg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab284388 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• 

  Western blot - Anti-alpha 1 Fetoprotein antibody [RM1011] (ab284388)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  

  Negative control: MDA-MB-231.

  Exposure time: Lane 4-5: 26 seconds ; Others: 5 seconds

  All lanes: Western blot - Anti-alpha 1 Fetoprotein antibody [RM1011] (ab284388) at 1/1000 dilution

  Lane 1: HepG2 (Human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: Huh7 (Human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 4: Hepa1-6 (Mouse hepatoma epithelial cell) whole cell lysate at 20 µg

  Lane 5: Hepa1-6 treated with 300ng/ml BFA for 24h whole cell lysate at 20 µg

  Lane 6: Mouse placenta lysate at 20 µg

  Lane 7: Rat placenta lysate at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 68 kDa

  Observed band size: 68 kDa

• 

  Immunoprecipitation - Anti-alpha 1 Fetoprotein antibody [RM1011] (ab284388)

  alpha 1 Fetoprotein was immunoprecipitated from 0.35 mg HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 10ug with ab284388 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab284388 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  

  Lane 1: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 10ug

  Lane 2: abab284388 IP in HepG2 whole cell lysate

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab284388 in HepG2 whole cell lysate

  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  Exposure time: 3 seconds

  All lanes: Immunoprecipitation - Anti-alpha 1 Fetoprotein antibody [RM1011] (ab284388)

  Predicted band size: 68 kDa

  Observed band size: 68 kDa

• 

  Flow Cytometry - Anti-alpha 1 Fetoprotein antibody [RM1011] (ab284388)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell) cells labelling alpha 1 Fetoprotein with ab284388 at 1/500 dilution Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.

• 

  Indirect ELISA - Anti-alpha 1 Fetoprotein antibody [RM1011] (ab284388)

  ELISA using ab284388 at varying antibody concentrations and antigen concentration at 1000 ng/ml. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody.