Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Chromogranin A.
(Peptide available as ab93753.)
Our Abpromise guarantee covers the use of ab85554 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
アプリケーション | Abreviews | 特記事項 |
---|---|---|
IHC-P | Use a concentration of 5 µg/ml. | |
WB | Use a concentration of 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa). | |
IP | Use a concentration of 5 µg/ml. | |
ICC/IF | Use a concentration of 1 - 5 µg/ml. |
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
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.
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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