Anti-SNAP25 antibody [EPR3275] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-SNAP25 antibody [EPR3275] - BSA and Azide free (AB171355)

Flow Cytometry analysis of Neuro-2a(Mouse neuroblastoma neuroblast) cells labelling SNAP25 with Purified ab109105 at 1 : 200 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor™ 488, ab150077) secondary antibody was used at 1 : 2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using ab109105, the same antibody clone in a different buffer formulation.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SNAP25 antibody [EPR3275] - BSA and Azide free (AB171355)

Immunocytochemistry analysis of Neuro-2a(Mouse neuroblastoma neuroblast) cells labeling SNAP25 with Purified ab109105 at 1/250 dilution (8.9 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

This data was developed using ab109105, the same antibody clone in a different buffer formulation.

• WB

Lab

Western blot - Anti-SNAP25 antibody [EPR3275] - BSA and Azide free (AB171355)

False colour image of Western blot : Anti-SNAP25 antibody [EPR3275] staining at 1/1000 dilution, shown in green; loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution, shown in red. In Western blot, ab109105 was shown to bind specifically to SNAP25. A band was observed at 27 kDa in wild-type SH-SY5Y cell lysates with no signal observed at this size in SNAP25 knockout cell line ab280041 (knockout cell lysate ab280100). To generate this image, wild-type and SNAP25 knockout SH-SY5Y cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

This data was developed using ab109105, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-SNAP25 antibody [EPR3275] (<a href='/en-us/products/primary-antibodies/snap25-antibody-epr3275-ab109105'>ab109105</a>) at 1/1000 dilution

Lane 1:

Wild-type SH-SY5Y cell lysate at 20 µg

Lane 2:

SNAP25 knockout SH-SY5Y cell lysate at 20 µg

Lane 2:

Western blot - Human SNAP25 knockout SH-SY5Y cell line (<a href='/en-us/products/cell-lines/human-snap25-knockout-sh-sy5y-cell-line-ab280041'>ab280041</a>)

Lane 3:

Neuro-2a cell lysate at 20 µg

Lane 4:

K562 cell lysate at 20 µg

Predicted band size: 23 kDa

Observed band size: 27 kDa

false

• WB

Unknown

Western blot - Anti-SNAP25 antibody [EPR3275] - BSA and Azide free (AB171355)

This data was developed using ab109105, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-SNAP25 antibody [EPR3275] (<a href='/en-us/products/primary-antibodies/snap25-antibody-epr3275-ab109105'>ab109105</a>) at 1/1000 dilution

Lane 1:

Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate

Lane 2:

Mouse brain lysate

Lane 3:

Mouse hippocampus lysate

Lane 4:

Rat brain lysate

Lane 5:

Rat hippocampus lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 23 kDa

Observed band size: 25 kDa

false

• WB

Unknown

Western blot - Anti-SNAP25 antibody [EPR3275] - BSA and Azide free (AB171355)

This data was developed using ab109105, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-SNAP25 antibody [EPR3275] (<a href='/en-us/products/primary-antibodies/snap25-antibody-epr3275-ab109105'>ab109105</a>) at 1/1000 dilution

All lanes:

Human brain lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 23 kDa

Observed band size: 25 kDa

false