Anti-SNAP25 抗体 [EPR3275]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SNAP25 antibody [EPR3275] (AB109105)

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.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-SNAP25 antibody [EPR3275] (AB109105)

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).

• WB

Lab

Western blot - Anti-SNAP25 antibody [EPR3275] (AB109105)

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.

All lanes:

Western blot - Anti-SNAP25 antibody [EPR3275] (ab109105) 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='/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

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• WB

Supplier Data

Western blot - Anti-SNAP25 antibody [EPR3275] (AB109105)

All lanes:

Western blot - Anti-SNAP25 antibody [EPR3275] (ab109105) at 1/1000 dilution

All lanes:

Human brain lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/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

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• WB

Lab

Western blot - Anti-SNAP25 antibody [EPR3275] (AB109105)

All lanes:

Western blot - Anti-SNAP25 antibody [EPR3275] (ab109105) at 1/1000 dilution

Lane 1:

Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 2:

Mouse brain lysate at 20 µg

Lane 3:

Mouse hippocampus lysate at 20 µg

Lane 4:

Rat brain lysate at 20 µg

Lane 5:

Rat hippocampus lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/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

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• WB

CiteAb

Western blot - Anti-SNAP25 antibody [EPR3275] (AB109105)

SNAP25 western blot using anti-SNAP25 antibody [EPR3275] ab109105. Publication image and figure legend from Cui, Z., Zhang, Y., et al., 2018, Nat Commun, PubMed 30341298.

ab109105 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 ab109105 please see the product overview.

Screening of potent NAPIs in HepG2. a Immunoblots for autophagy-related proteins LC3-II, p62 (left); semi-quantified analysis (n = 3) in various nanoparticles treated cells (right). CQ and Rapamycin (Rapa) were used as positive controls for autophagy inhibition and autophagy activation, respectively. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the loading control. Normalized band densities were shown below each band. b Fluorescence images of mCherry-GFP-LC3 cells after incubation with CQ or NDs for 48 h (autophagosomes : mCherry+/GFP+ yellow puncta; autolysosomes : mCherry+/GFP) and quantification of the number of LC3 puncta per cell in cells (10 cells per group). Scale bar : 10 μm. c Immunoblots for autophagy-related proteins LC3-II, p62 (left); semi-quantified analysis (n = 3) in CQ, NDs, or CQ–NDs-treated cells (right). &p < 0.05, significantly different from NDs. GAPDH was used as the loading control. Normalized band densities were shown below each band. d Left : Cell viability after incubation with ATO or various NAPIs–ATO mixture for 48 h (n = 3). ##p < 0.01 by t-test, significantly different from ATO. Right : Cell viability after 48 h NDs–ATO treatment with RNAi of autophagy proteins ATG5 and ATG7 (n = 3). e, f Immunoblots for autophagy-related protein LC3-II and autolysosomal process-related protein NUPR1, SNAP25, VAMP8 in NDs-treated cells. g Immunoblots for autolysosomal process-related protein NUPR1 after NDs treatment with RNAi of autophagy proteins ATG5 and ATG7. GAPDH was used as the loading control. Error bars are s.d.

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