Anti-SNAP25 antibody

• WB

Project

Western blot - Anti-SNAP25 antibody (AB41455)

All lanes:

Western blot - Anti-SNAP25 antibody (ab41455) at 1 µg/mL

Lane 1:

Spinal cord (Rat) tissue lysate at 10 µg

Lane 2:

Hippocampus (Mouse) tissue Lysate at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 23 kDa

Observed band size: 26 kDa

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• ICC

Lab

Immunocytochemistry - Anti-SNAP25 antibody (AB41455)

ab41455 staining SNAP25 in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab41455 at 1μg/ml and ab192757, Mouse mono Anti-PSD95 antibody [K28/43] - Synaptic Marker. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC

Lab

Immunocytochemistry - Anti-SNAP25 antibody (AB41455)

ab4455 staining SNAP25 in PC12 cells. The cells were fixed with 100% methanol (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab41455 at 5μg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 μM for 1hour at room temperature.

• IP

Unknown

Immunoprecipitation - Anti-SNAP25 antibody (AB41455)

SNAP25 was immunoprecipitated using 0.5mg Rat Spinal Cord tissue lysate, 5µg of Rabbit polyclonal to SNAP25 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Rat Spinal Cord tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab41455.
Secondary : Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
Band : 26kDa; SNAP25

All lanes:

Immunoprecipitation - Anti-SNAP25 antibody (ab41455)

Predicted band size: 23 kDa

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• WB

Unknown

Western blot - Anti-SNAP25 antibody (AB41455)

All lanes:

Western blot - Anti-SNAP25 antibody (ab41455) at 1 µg/mL

Lane 1:

Marker

Lane 2:

Zebrafish brain homogenate at 20 µg

Lane 3:

SH-SY5Y (Human neuroblastoma cell line) whole cell lysate at 20 µg

Lane 4:

Mouse brain homogenate at 20 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution

Predicted band size: 23 kDa

Observed band size: 26 kDa

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Exposure time: 2min

• WB

Unknown

Western blot - Anti-SNAP25 antibody (AB41455)

All lanes:

Western blot - Anti-SNAP25 antibody (ab41455) at 1 µg/mL

All lanes:

Western blot - Recombinant Human SNAP25 protein (Tag Free) (<a href='/en-us/products/proteins-peptides/recombinant-human-snap25-protein-ab74529'>ab74529</a>) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 23 kDa

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Exposure time: 1min

• WB

CiteAb

Western blot - Anti-SNAP25 antibody (AB41455)

SNAP25 western blot using anti-SNAP25 antibody ab41455. Publication image and figure legend from Bray, J. K., Elgamal, O. A., et al., 2020, Cell Death Dis, PubMed 32080178.

ab41455 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 ab41455 please see the product overview.

Transgenic mouse model with conditional loss of REST in the exocrine pancreas.a Schematic of REST knockout breeding. A breeder with Cre Recombinase gene downstream of the p48 promotor is crossed with a breeder containing LoxP sites (red triangles) flanking exon two of REST. White box indicates non-coding UTR. Yellow indicates zinc fingers. Gray "P" arrows indicate target region of PCR primers. b Gel electrophoresis of PCR products from genomic DNA for Cre Recombinase (P1 and P2) (~450 bps) and floxed Rest (P3 and P4) (WT : ~550 bps; Fl : ~700 bps). c Gel electrophoresis of PCR products from primers P5 and P4 encompassing LoxP sites of Rest exon two from pancreatic genomic DNA of 1–2-month-old mice. Wild-type product is ~2.8 kbs, floxed exon two product is ~2.9 kbs, and successfully excised exon two product is ~1.1 kb (n = 3) (Primer designs30). d Sanger Sequencing of PCR product (P4 and P5) from Cre/RESTfl/fl pancreatic DNA. Black indicates Rest introns, gray indicates original vector backbone, and red indicates LoxP sequence (n = 3). e qRT-PCR of Rest exon gene expression of 6-week-old mouse pancreas (normalized to 18S rRNA) (n = 3–4) (Avg ± SD). f Western blot of pancreatic protein with anti-Rest antibody (Proteintech) (MW = 200 kDa) (n = 3). g Western blot of pancreatic protein for Rest target gene Snap25 (MW = 30 kDa) (n = 3). h Gene expression of six-week-old Cre/RESTfl/fl pancreas relative to p48-Cre (normalized to 18S rRNA) (n = 4) (Avg ± SD). *p < 0.05, **p < 0.001, ***p < 0.0001.

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