Anti-Chromogranin A抗体(ab45179)

Unpurified ab45179 staining Chromogranin A in pig small intestine tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/750 in blocking buffer) for 2 hours at 21°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.

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Blocking and diluting buffer: 5% NFDM/TBST.

ab45179 staining Chromogranin A in SHSY5Y cells. The cells were fixed with 100% methanol (5 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 ab45179 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 ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) 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.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human adenocarcinoma of pancreas tissue labelling Chromogranin A with purified ab45179 at 1/200. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

Blocking and diluting buffer: 5% NFDM/TBST.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse pancreas tissue labelling Chromogranin A with purified ab45179 at 1/200. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

Chromogranin A staining on monkey pancreas sections. The sections were subjected to heat mediated epitope retrieval using citric acid (pH 6) and blocked using 1% BSA for 10 mins at 21°C. Unpurifeid ab45179 was used at a dilution of 1/750 using TBS/BSA/Azide for 2 hours at 21°C. (Red arrowheads indicate extra Islet positive cells).

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