Anti-Synaptophysin antibody [EP1098Y] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Synaptophysin antibody [EP1098Y] - BSA and Azide free (AB214621)

Overlay histogram showing PC12 cells stained with ab52636 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52636, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (0.5μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in PC12 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions. Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52636).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Synaptophysin antibody [EP1098Y] - BSA and Azide free (AB214621)

This ICC data was generated using the same anti-Synapthophysin antibody clone, EP1098Y, in a different buffer formulation (cat# ab52636).

PC12 cells labelled with ab52636 at 1/250 - 1/500 dilution

• WB

Lab

Western blot - Anti-Synaptophysin antibody [EP1098Y] - BSA and Azide free (AB214621)

This data was developed using the same antibody clone in a different buffer formulation (ab52636).

Lanes 1-3 : Merged signal (red and green). Green - ab52636 observed at 38 kDa. Red - loading control, ab7291 observed at 50 kDa.

ab52636 Anti-Synaptophysin antibody [EP1098Y] was shown to specifically react with Synaptophysin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267272 (knockout cell lysate ab257060) was used. Wild-type and Synaptophysin knockout samples were subjected to SDS-PAGE. ab52636 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti- Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti- Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Synaptophysin antibody [EP1098Y] - Synaptic Marker (<a href='/en-us/products/primary-antibodies/synaptophysin-antibody-ep1098y-synaptic-marker-ab52636'>ab52636</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

SYP knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human SYP (Synaptophysin) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-syp-synaptophysin-knockout-hek-293t-cell-line-ab267272'>ab267272</a>)

Lane 3:

Human brain tissue lysate

Predicted band size: 33 kDa

Observed band size: 38 kDa

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