Anti-Chromogranin A 抗体 (ab85554)

Anti-Chromogranin A antibody 画像

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  Immunoprecipitation - Anti-Chromogranin A antibody (ab85554)

  Chromogranin A was immunoprecipitated using 0.5mg Mouse Pancreas tissue lysate, 5µg of Rabbit polyclonal to 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 Pancreas tissue lysate 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°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab85554.

  Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

  Band: 50kDa; Chromogranin A

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  Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody (ab85554)

  ICC/IF image of ab85554 stained PANC-1 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 ab85554 at 5 µg/mL overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) used at a 1/250 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.

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  Western blot - Chromogranin A antibody (ab85554)
  Anti-Chromogranin A antibody (ab85554) at 1 µg/ml + Human pancreas tissue lysate - total protein (ab29816) at 10 µg
  
  Secondary
  Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
  Developed using the ECL technique
  
  Performed under reducing conditions.
  
  Predicted band size : 50 kDa
  Observed band size : 50 kDa
  Additional bands at : 72 kDa. We are unsure as to the identity of these extra bands.
  
  Exposure time : 20 minutes
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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Chromogranin A antibody (ab85554)
  IHC image of Chromogranin A staining in Human Pancreas Adenocarcinoma FFPE section, performed on a BondTM system using the standard protocol F. 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 ab85554, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX
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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody (ab85554)This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United Kingdom

  IHC image of Chromogranin A staining in Monkey Pancreas sections, performed The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6). Sections were subjected to 2% H₂0₂O then Avidin-Biotin to block endogenous peroxidases/biotin. The section was then incubated with ab85554, 1/5000, for 2 hours at 12°C. The section was then counterstained with haematoxylin

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