Anti-Heme Oxygenase 1 antibody [EP1391Y] - BSA and Azide free

8 product images

• 

  Western blot - Anti-Heme Oxygenase 1 antibody [EP1391Y] - BSA and Azide free (ab219360)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947).
  

  Lanes 1 - 7: Merged signal (red and green). Green - Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947 observed at 33 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

  Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947 was shown to react with Heme Oxygenase 1 in wild-type A549 cells in Western blot with loss of signal observed in HMOX1 knockout cell line Human HMOX1 (Heme Oxygenase 1) knockout A549 cell line ab269503 (knockout cell lysate Human HMOX1 (Heme Oxygenase 1) knockout A549 cell lysate ab269665). Wild-type A549 and HMOX1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  All lanes: Western blot - Anti-Heme Oxygenase 1 antibody [EP1391Y] (Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947) at 1/2000 dilution

  Lane 1: Wild-type A549 cell lysate at 20 µg

  Lane 2: HMOX1 knockout A549 cell lysate at 20 µg

  Lane 2: Western blot - Human HMOX1 (Heme Oxygenase 1) knockout A549 cell line (Human HMOX1 (Heme Oxygenase 1) knockout A549 cell line ab269503)

  Lane 3: Human Spleen tissue lysate at 20 µg

  Lane 4: HL-60 cell lysate at 20 µg

  Lane 5: MCF7 cell lysate at 20 µg

  Lane 6: HeLa cell lysate at 20 µg

  Lane 7: A549 cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 33 kDa

  Observed band size: 33 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heme Oxygenase 1 antibody [EP1391Y] - BSA and Azide free (ab219360)

  IHC image of Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947 staining in mouse spleen formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947, 5μg/ml, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heme Oxygenase 1 antibody [EP1391Y] - BSA and Azide free (ab219360)

  This data was developed using Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947, the same antibody clone in a different buffer formulation.
  

  Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Heme Oxygenase 1 with Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947 at 1/2000 (0.246 ug/ml) followed by a LeicaDS9800 (Bond Polymer Refine Detection) was used at Ready to use dilution. Cytoplasmic staining on Kupffer cells in human liver. The section was incubated with Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution.

  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-Heme Oxygenase 1 antibody [EP1391Y] - BSA and Azide free (ab219360)

  This data was developed using Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947, the same antibody clone in a different buffer formulation.
  

  Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Heme Oxygenase 1 with Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947 at 1/2000 (0.246 ug/ml) followed by a LeicaDS9800 (Bond Polymer Refine Detection) was used at Ready to use dilution. Cytoplasmic staining on mouse spleen. The section was incubated with Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution.

  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-Heme Oxygenase 1 antibody [EP1391Y] - BSA and Azide free (ab219360)

  This data was developed using Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947, the same antibody clone in a different buffer formulation.
  

  Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Heme Oxygenase 1 with Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947 at 1/2000 (0.246 ug/ml) followed by a LeicaDS9800 (Bond Polymer Refine Detection) was used at Ready to use dilution. Cytoplasmic staining on human spleen. The section was incubated with Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947 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 LeicaDS9800 (Bond Polymer Refine Detection) was used at Ready to use dilution.

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• 

  Immunoprecipitation - Anti-Heme Oxygenase 1 antibody [EP1391Y] - BSA and Azide free (ab219360)

  Heme Oxygenase 1 was immunoprecipitated from 0.35mg mouse spleen lysate with Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947 at 1/1000 dilution (1 μg/mL). VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used as the secondary antibody at 1/1000 dilution.
  

  Lane 1: Mouse spleen tissue lysate 10 μg
  Lane 2: Mouse spleen tissue lysate
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947 in mouse spleen lysate.

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

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947).

  All lanes: Immunoprecipitation - Anti-Heme Oxygenase 1 antibody [EP1391Y] (Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947)

  Predicted band size: 33 kDa

  Observed band size: 33 kDa

  Exposure time: 3s

• 

  Flow Cytometry (Intracellular) - Anti-Heme Oxygenase 1 antibody [EP1391Y] - BSA and Azide free (ab219360)

  Overlay histogram showing HEK293 cells stained with Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HEK293 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947).
  

• 

  Flow Cytometry (Intracellular) - Anti-Heme Oxygenase 1 antibody [EP1391Y] - BSA and Azide free (ab219360)

  This data was developed using Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947, the same antibody clone in a different buffer formulation.
  

  Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling Heme Oxygenase 1 with Anti-Heme Oxygenase 1 antibody [EP1391Y] ab52947 at 1/50 dilution (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) ab150077) at 1/2000 dilution was used as the secondary antibody.