Anti-SNAP25 antibody [EP3274]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SNAP25 antibody [EP3274] (AB108990)

ab108990 showing negative staining in human Normal colon tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SNAP25 antibody [EP3274] (AB108990)

Immunohistochemical analysis of paraffin-embedded human Alzheimer's brain tissue* labelling SNAP25 with ab108990 at 1ug/ml followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Positive staining in human Alzheimer's brain.

The section was incubated with ab108990 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0 epitope retrieval solution1) for 20 mins.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SNAP25 antibody [EP3274] (AB108990)

ab108990, at 1/100 dilution, staining SNAP25 in paraffin-embedded Human cerebellum tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SNAP25 antibody [EP3274] (AB108990)

ab108990, at 1/100 dilution, staining SNAP25 in paraffin-embedded Human brain tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SNAP25 antibody [EP3274] (AB108990)

ab108990 showing negative staining in human Skeletal muscle tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SNAP25 antibody [EP3274] (AB108990)

ab108990 showing negative staining in human Normal spleen tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-SNAP25 antibody [EP3274] (AB108990)

False colour image of Western blot : Anti-SNAP25 antibody [EP3274] 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, ab108990 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.

All lanes:

Western blot - Anti-SNAP25 antibody [EP3274] (ab108990) 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 [EP3274] (AB108990)

All lanes:

Western blot - Anti-SNAP25 antibody [EP3274] (ab108990) at 1/1000 dilution

Lane 1:

Fetal brain lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

SH-SY5Y cell lysate at 10 µg

Lane 4:

U87-MG cell lysate at 10 µg

Lane 5:

Fetal hippocampus lysate at 10 µg

Lane 6:

Fetal cerebellum lysate at 10 µg

Lane 7:

Neuro-2a cell lysate at 10 µg

Predicted band size: 23 kDa

false

• WB

Unknown

Western blot - Anti-SNAP25 antibody [EP3274] (AB108990)

Blocking/Diluting buffer and concentration : 5% NFDM /TBST

All lanes:

Western blot - Anti-SNAP25 antibody [EP3274] (ab108990) at 1/2000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Mouse spleen tissue lysate at 20 µg

Lane 5:

Rat brain tissue lysate at 20 µg

Lane 6:

Rat heart tissue lysate at 20 µg

Lane 7:

Rat kidney tissue lysate at 20 µg

Lane 8:

Rat spleen tissue lysate at 20 µg

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>)

Predicted band size: 23 kDa

Observed band size: 25 kDa

false

• WB

CiteAb

Western blot - Anti-SNAP25 antibody [EP3274] (AB108990)

SNAP25 western blot using anti-SNAP25 antibody [EP3274] ab108990. Publication image and figure legend from Almandoz-Gil, L., Persson, E., et al., 2018, Front Neurol, PubMed 29623065.

ab108990 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab108990 please see the product overview.

Representative images of immunofluorescence staining of non-tg cortical primary neurons (12 DIV) using antibodies against alpha-synuclein (αSyn) [mAb1338, red, (A)], VAMP-2 [(green), (B)], SNAP-25 [(green), (C)], and syntaxin-1 [(green), (D)]. DAPI in blue. Scale bar 50 µm.

false