On D4, D5, and D6, the cells were washed with ice-cold PBS (Invitrogen). Cell lysates were prepared using RIPA buffer, as previously described [30 (link)]. Equal volumes of cell lysates were applied to SDS-PAGE, and transferred to PVDF for immunoblotting [30 (link)]. The following antibodies were used at 1/1000 dilution: anti-synaptophysin (Cell Signaling 4329S), anti-SNAP25 (Santa Cruz sc-7538), anti-PSD95 (Cell Signaling, 2507S), anti-GAP43 (Cell signaling, 8945S), and anti-β-tubulin (Sigma T5201). Secondary antibodies, anti-mouse HRP-linked, and anti-rabbit HRP linked from Cell Signaling and anti-goat HRP-linked from Santa Cruz were used to detect the bands with Supersignal chemiluminescence ECL kit (Millipore, Molsheim, France). Gel images were obtained while using the Fusion Imaging System (Fusion Fx5; Vilber Lourmat, France), and densitometric analysis was performed using the freeware ImageJ.
Anti snap25
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Anti-SNAP25 is a primary antibody that targets the SNAP25 protein, which is a key component of the SNARE complex involved in synaptic vesicle fusion and neurotransmitter release. This antibody can be used for the detection and analysis of SNAP25 in various applications, such as Western blotting, immunohistochemistry, and immunocytochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Anti psd95, by Cell Signaling Technology (1 mentions)
Supersignal chemiluminescence ecl kit, by Merck Group (1 mentions)
Anti β tubulin, by Merck Group (1 mentions)
Anti synaptophysin, by Cell Signaling Technology (1 mentions)
Anti gap43, by Cell Signaling Technology (1 mentions)
Anti mouse hrp linked, by Cell Signaling Technology (1 mentions)
Anti pimt, by Abcam (1 mentions)
Anti mafa, by Cell Signaling Technology (1 mentions)
Anti hdac5, by Cell Signaling Technology (1 mentions)
Anti gapdh, by Cell Signaling Technology (1 mentions)
Anti pdx1, by Cell Signaling Technology (1 mentions)
Anti insulin, by Cell Signaling Technology (1 mentions)
Anti neurod1, by Cell Signaling Technology (1 mentions)
Anti syp, by Santa Cruz Biotechnology (1 mentions)
Ab20066, by Abcam (1 mentions)
Ab85367, by Abcam (1 mentions)
Ab111468, by Abcam (1 mentions)
Anti munc18, by Synaptic Systems (1 mentions)
Sc 13937, by Santa Cruz Biotechnology (1 mentions)
3 protocols using anti snap25
1
Quantitative Immunoblotting of Synaptic Proteins
2
Protein Expression Analysis in BRIN-BD11 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BRIN-BD11 cells were lysed using TENNS lysis buffer [120 mM of NaCl, 1% Triton X-100, 20 mM of Tris–HCl pH 7.5, 10% glycerol, 2 mM of EDTA and protease inhibitor cocktail (10 μg/mL each of aprotinin and leupeptin)]. Approximately 30 μg of protein was resolved by 10% SDS-PAGE and a Western blot analysis was carried out using specified antibodies. The catalogue and primary dilution of the antibodies used in the study are Anti-PIMT (Abcam, Cambridge, UK, catalogue no. Ab70559) 1:2000; anti-PDX1 (Cell Signaling Technologies, Danvers, MA, USA, catalogue no. 5679) 1:1000; anti-MafA (sc-390491) 1:200); anti-HDAC5 (sc-133225) 1:1000; anti-GCK (sc-17819) 1:200; anti-insulin (Cell Signaling Technologies, 8138) 1:1000; anti-Synaptotagmin13 (Novus Biologicals, NB2-20546) 1:1000; anti-SNAP25 (sc-390644 Santa Cruz Biotechnology, Finnell Street Dallas, TX, USA) 1:200; anti-Kir6.2 (sc-20809) 1:200; anti-NeuroD1 (Cell Signaling Technologies, 2833) 1:1000; and anti-GAPDH (Cell Signaling Technologies, 2118) 1:5000. The protein levels were normalized using GAPDH and Western blots were quantified by using ImageJ [32 (link)].
3
Western Blot Analysis of Neurotransmitter Receptors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunoblot analysis was carried out under standard conditions (Marrocco et al., 2012 (link)) using the following primary antibodies: anti-tyrosine hydroxylase (TH H126, Santa Cruz, 1:1000); anti-D1 receptors (Ab20066, Abcam, 1:500), anti-D2 receptors (Ab85367, Abcam, 1:500), anti-DAT (high affinity DA transporter) (ab111468, Abcam, 1:1000); anti-adenosine receptors (sc-13937, Santa Cruz, 1:500), synaptic vesicle-associated proteins: anti-Rab3a (#107111, Synaptic Systems, 1:2000); anti-Munc18 (#116011, Synaptic Systems 1:2000; anti-SNAP 25 (sc-136,267, Santa Cruz 1:5000); anti-SYP (sc-9116 Santa Cruz 1:8000), and anti-syntaxin (sc-13994, Santa Cruz 1:4000). Secondary antibodies directed against rabbit or mouse antibodies (Amersham) were used at 1:7500 dilution. After immunoblotting, digitized images of bands immunoreactive to the target antibodies and actin were acquired (FUSION®) and the area of immunoreactivity corresponding to each band was measured using ImageJ imaging software. The ratios of the targets to actin were then determined and these values were compared to check statistical significance.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!