Anti-ALDH1L1 antibody [3E9] - Astrocyte Marker

5 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALDH1L1 antibody [3E9] (ab56777)

  ALDH1L1 antibody (ab56777) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human kidney.
  

  This image was generated using the ascites version of the product.

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  Flow Cytometry - Anti-ALDH1L1 antibody [3E9] - Astrocyte Marker (ab56777)

  ALDH1L1 Flow Cytometry staining using mouse Anti-ALDH1L1 antibody

  Overlay histogram showing HepG2 cells stained with ab56777 (red line). The cells were fixed with 80% methanol (5 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 (ab56777, 0.5μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was Mouse IgG3 [MG3-35] (Mouse IgG3, kappa monoclonal [MG3-35] - Isotype Control ab18394, 1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
  
  

  This image was generated using the ascites version of the product.

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  Western blot - Anti-ALDH1L1 antibody [3E9] - Astrocyte Marker (ab56777)

  ALDH1L1 Western blot staining using mouse Anti-ALDH1L1 antibody

  ALDH1L1 antibody (ab56777) at 1ug/lane + NIH/3T3 cell lysate at 25 μg/lane.
  

  This image was generated using the ascites version of the product.

  All lanes: Western blot - Anti-ALDH1L1 antibody [3E9] - Astrocyte Marker (ab56777)

  Predicted band size: 99 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-ALDH1L1 antibody [3E9] (ab56777)

  ICC/IF image of ab56777 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab56777, 1μg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.
  
  

  This image was generated using the ascites version of the product.

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  Immunoprecipitation - Anti-ALDH1L1 antibody [3E9] - Astrocyte Marker (ab56777)

  ALDH1L1 was immunoprecipitated using 0.5mg Mouse Liver whole tissue extract, 5μg of Mouse monoclonal to ALDH1L1 and 50μl of protein G magnetic beads (+). No antibody was added to the control (-).
  The antibody was incubated under agitation with Protein G beads for 10min, Mouse Liver whole tissue extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
  Proteins were eluted by addition of 40μl SDS loading buffer and incubated for 10min at 70^oC; 10μl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab56777.
  Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
  Band: 99kDa: ALDH1L1; non specific - 60kDa: We are unsure as to the identity of this extra band.
  

  This image was generated using the ascites version of the product.

  All lanes: Immunoprecipitation - Anti-ALDH1L1 antibody [3E9] - Astrocyte Marker (ab56777)

  Predicted band size: 99 kDa