The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 122,128 kDa (predicted molecular weight: 128 kDa).
1/1000 - 1/3000.
Tomosyn plays a regulatory role in calcium-dependent exocytosis and neurotransmitter release by inhibiting the assembly of trans-SNARE complexes between transport vesicles and the plasma membrane, via the displacement of munc18 from syntaxin-1. There are three splice variants of tomosyn designated m-tomosyn, b-tomosyn and s-tomosyn. Although b-tomosyn is ubiquitously expressed, s-tomosyn and m-tomosyn are expressed primarily in brain.
Cell Membrane and Cytoplasmic.
Note: Cytoplasmic, and associated with vesicular membranes and the plasma membrane. Detected at synapses and on synaptic vesicles
Anti-Tomosyn antibody (ab41271) at 1 µg/ml + Brain (Rat) Tissue Lysate - normal tissue at 10 µg
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 128 kDa Observed band size: 128 kDa Additional bands at: 40 kDa. We are unsure as to the identity of these extra bands.
Tamosyn is phosphorylated on a number of serine and threonine residues (SwissProt), which may account for the banding pattern observed. ab41271 is targeted against all isoforms of the Tamosyn protein.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Tomosyn antibody (ab41271)This image is courtesy of an abreview submitted by Sophie Pezet, CNRS, Paris, France
Immunohistochemistical detection of tomosyn using antibody (ab41271) on PFA perfusion fixed, free floating rat brain sections. Primary Antibody ab41271 was diluted 1/3000 and incubated for 18 hours @ 20°C in PBS + 0.3 % Triton X100. Secondary antibody: goat anti-rabbit Alexa Fluor® 488 (1/1000). The antibody produced a diffuse staining in many brain areas, corresponding to the staining published by Fujita 1998 (Nature). The submitted image is of hippocampal staining.