Anti-Synaptophysin antibody [SY38] (ab8049)

ab8049 staining Synaptophysin in Mouse brain tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citric acid. Samples were incubated with primary antibody (1/100) for 16 hours at 21°C. A Biotin-conjugated Goat anti-mouse IgG polyclonal (1/300) was used as the secondary antibody.

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All lanes : Anti-Synaptophysin antibody [SY38] (ab8049) at 1/500 dilution

Lane 1 : Human brain tissue lysate - total protein (ab29466)
Lane 2 : Brain (Mouse) Tissue Lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed at 1/3000 dilution

Predicted band size: 34 kDa
Observed band size: 38 kDa (why is the actual band size different from the predicted?)

ab8049 staining Synaptophysin in mouse hippocampus tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with formaldehyde and blocked with 5% BSA for 1 hour at 24°C. Samples were incubated with primary antibody (1/100 in BSA) for 12 hours at 4°C. An Alexa Fluor^® 488-conjugated donkey anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.

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ab8049 staining MAFA in human colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in citric acid. Samples were incubated with primary antibody (1/100 in TBS/BSA/azide) for 16 hours at 21°C. A Biotin-conjugated goat anti-mouseIgG polyclonal (1/300) was used as the secondary antibody.

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The pictures show immunohistochemical staining of normal mucosa and neuroendocrine tumour of the small intestine.  The antibody (ab8049) stains positive in a 1:10 dilution in nerve cells of the colon as well as some cells in normal colon mucosa and some neuroendocrine tumors of the digestive tract. Staining protocol: Mouse monoclonal to Synaptophysin (ab8049), diluted 1:10 in PBS plus BSA, 60 min at room temperature,  
secondary antibody: HRP solution with anti mouse polyclonal antibodies, 30min. DAB staining 10 min; Hematoxylin 30 sec.

These images were kindly supplied as part of the review submitted by Karin Birkenkamp-Demtroeder.

Overlay histogram showing PC12 cells stained with ab8049 (red line). The cells were fixed with 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 (ab8049, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in PC12 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.