Technical Data
S9108-19
Synaptophysin
Description:
Synaptophysin is an ~38kD integral membrane protein involved in neurotransmitter exocytosis present on synaptic vesicles (1). This protein consists of four transmembrane domains, with its amino-and carboxyl-terminus facing the cytoplasm. Due to its transmembrane domain structure, synaptophysin has been postulated to be a gap junction-like protein as voltage-dependent channel activity is seen when synaptophysin hexamers are reconstituted into lipid membranes (2,3). Recent studies have shown synaptophysin to be a major cholesterol-binding protein in brain synaptic vesicles (4) and it has been proposed that this cholesterol-binding ability may be of key importance in the generation of synaptic vesicles at plasma membranes (5). A regional reduction of synaptophysin was found in patients with Alzheimers disease (7) and schizophrenia (6).

Applications: Suitable for use in Western Blot, Immunohistochemistry. Other applications not tested.

Recommended Dilution:
Immunohistology: (Formalin/paraffin) 1:50 for 30 min at RT. Excellent for staining of formalin/paraffin with no special pretreatment.
With pretreatment: 1:200 for 30 min. No pretreatment is necessary, however staining of formalin fixed tissues is enhanced by boiling tissue sections in 10mM citrate buffer, pH 6 for 10-20 min followed by cooling at RT for 20 min. In combination with anti-chromogranin A and anti-NSE, antibody to synaptophysin is very useful in the identification of normal neuroendocrine cells and neuroendocrine neoplasms.

Optimal dilutions to be determined by the researcher.

Epitope: C-terminus

Positive Control:
Pancreas or neuroectodermal and neuroendocrine tumors

Cellular Localization:
Cytoplasmic

Storage and Stability:
May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Store at -20°C or colder. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Rabbit MAbs:
Monoclonal antibodies have traditionally been produced in a mouse. Why a mouse? Because, as a host, it is cost effective, easy to manage, and truly a standard in many biotechnology applications. But manufacturers and technicians alike have seen the limitations of mouse monoclonals, in that the resulting antibodies can have low affinities, which means that they have a weak attraction to bind to the protein targets. At times, the affinities are so low that extreme pretreatments, such as boiling the tissue sections, must be employed to modify the target, so that the low affinity antibody has a better chance to bind. Others have used special diluents to obtain better antibody-to-target binding. By modifying the pH and the salts in the buffers and/or diluents, the antibody can be modified such that it is in its optimal binding state. Like pretreaments, this too will only optimize an already low affinity antibody. Instead of modifying the target, or the diluent, to give a weak antibody a better chance to bind, why not produce a higher affinity antibody?

The concept itself is simple.  Antibodies produced by rabbits are known to have a higher affinity than antibodies raised in mice. Cloning techniques produce highly specific antibodies. Additionally, the rabbit immuno-response recognizes antigens (epitopes) that are not immunogenic in mice. Our technique combines these two production methods to produce… a RABBIT MONOCLONAL. The resulting rabbit monoclonal antibody has ten times the affinity of the mouse antibody, thus resulting in a more SPECIFIC and much more SENSITIVE antibody.  Studies with these Rabbit MAbs have shown that pretreaments may no longer be necessary to achieve optimal staining results, but may only be needed to standardize the tissue conditions. This technique will continue to be employed to produce other important markers, for Ki67, Cyclin D1, and CD3, antibodies that historically are difficult in immunohistochemical applications.
TypeIsotypeCloneGrade
MabIgG5G219Supernatant
SizeStorageShippingSourceHost
250ul-20°CBlue IceHumanRabbit
Concentration:
Not determined
Immunogen:
A synthetic peptide of human synaptophysin.
Purity:
Tissue culture supernatant
Form
Supplied as a liquid with 0.09% sodium azide.
Specificity:
Recognizes a protein of 38kD, identified as synaptophysin. It labels normal neuroendocrine cells of human adrenal medulla, carotid body, skin, pituitary gland, thyroid, lung, pancreas, gastrointestinal mucosa, Paneth’s cells in the gastrointestinal tract and of gastric parietal cells. Neurons in the brain, spinal cord, and retina are also labeled.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Honer, W.G., Hu, L. and Davies, P., Brain Research 609: 9-20 (1993). 2. Rehm, H., Wiedenmann, B. and Betz, H., EMBO J. 5: 535-541 (1986). 3. Thomas, L., et al., Science 242: 1050-1053 (1988). 4. Thiele, C., et al., Nat. Cell Biol. 2: 42-49 (2000). 5. Martin, T.F., Nat. Cell Biol. 2: E9-E11 (2000). 6. Landen, M., et al., Biol. Psychiatry 46: 1698-1702 (1999). 7. Honer, W., et al., Neurobiol. Aging 13: 373-382 (1992).