Anti-Chromogranin A antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody (AB85554)

ab85554 staining Chromogranin-A precursor in SHSY5Y cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab85554 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IP

Lab

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

All lanes:

Immunoprecipitation - Anti-Chromogranin A antibody (ab85554)

Predicted band size: 50 kDa

true

Exposure time: 20min

• IHC-P

AbReview19889****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody (AB85554)

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

This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United Kingdom

• WB

Unknown

Western blot - Anti-Chromogranin A antibody (AB85554)

All lanes:

Western blot - Anti-Chromogranin A antibody (ab85554) at 1 µg/mL

All lanes:

Human pancreas tissue lysate - total protein (<a href='/en-us/products/unavailable/human-pancreas-tissue-lysate-total-protein-ab29816'>ab29816</a>) at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 50 kDa

Observed band size: 50 kDa,72 kDa

true

Exposure time: 20min