Anti-Synapsin I antibody - Synaptic Marker

• WB

Project

Western blot - Anti-Synapsin I antibody - Synaptic Marker (AB64581)

The immunogen for ab64581 is predicted to recognise both the 70 kDa and 74 kDa isoforms of Synapsin I. Synapsin I is also known to contain a number of potential phosphorylation and glycosylation sites (SwissProt data).

All lanes:

Western blot - Anti-Synapsin I antibody - Synaptic Marker (ab64581) at 1 µg/mL

Lane 1:

Spinal Cord (Mouse) Tissue Lysate at 10 µg

Lane 2:

Brain (Mouse) Tissue Lysate at 10 µg

Lane 3:

Brain (Rat) Tissue Lysate - normal tissue at 10 µg

Secondary

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Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 74 kDa

Observed band size: 70 kDa,74 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-Synapsin I antibody - Synaptic Marker (AB64581)

Synapsin I - Synaptic Marker was immunoprecipitated using 0.5mg Mouse Brain whole tissue lysate, 5µg of Rabbit polyclonal to Synapsin I - Synaptic Marker 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, Mouse Brain whole 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 ab64581.
Secondary : Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band : 80kDa : Synapsin I - Synaptic Marker contains a number of potential glycosylation sites (SwissProt) which may explain the higher migrating band.

All lanes:

Immunoprecipitation - Anti-Synapsin I antibody - Synaptic Marker (ab64581)

Predicted band size: 74 kDa

false

• WB

AbReview49096****

Western blot - Anti-Synapsin I antibody - Synaptic Marker (AB64581)

Blocked with 5% milk for 1 hour at 25°C.

Sample was incubated with primary antibody for 18 hours at 4°C in TBS + 0.1% Tween.

All lanes:

Western blot - Anti-Synapsin I antibody - Synaptic Marker (ab64581) at 1/5000 dilution

All lanes:

Rat whole brain tissue lysate at 15 µg

Secondary

All lanes:

Donkey anti-Rabbit IgG at 1/5000 dilution

Predicted band size: 74 kDa

Observed band size: 80 kDa,85 kDa

true

Exposure time: 15s

This image is courtesy of an anonymous Abreview