Invitrogen

Synaptophysin Monoclonal Antibody (SY38)

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Synaptophysin Antibody in Immunohistochemistry (PFA fixed) (IHC (PFA)) — Immunohistochemistry was performed on human cerebellum and human breast tissue. Tissue was deparaffinized with xylene, followed by rehydration in sequential washes of 100% ethanol, 95% ethanol, 80% ethanol, and water. To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0) and heated for 20 min. Following antigen retrieval, tissues were blocked in a 10% goat serum (Product # 31872) in wash buffer solution for 30 minutes at room temperature and endogenous peroxidase activity quenched with Peroxidase Suppressor (Product # 35000). Tissue was then probed with a Synaptophysin mouse monoclonal antibody (Product # MA1-213) at a dilution of 1:200 in 10% goat serum in wash buffer for 1 hour at room temperature in a humidified chamber. Negative control tissue received no primary antibody. Tissues were washed extensively with PBST, and detection was performed using a SuperBoost™ goat anti-mouse Poly HRP secondary antibody reagent (Product # B40961) followed by colorimetric detection using DAB Quanto (Product # TA-125-QHDX). Tissues were then counterstained with hematoxylin and prepped for mounting and imaging.

Synaptophysin Antibody in Western Blot (WB) — Western blot analysis of Synaptophysin was performed by loading 20 µg of indicated whole cell lysates and 7 µL of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BX10). Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% Milk in TBST for 1 hour at room temperature. Synaptophysin was detected at ~38 kDa using a mouse monoclonal antibody (Product # MA1-213, upper) at a dilution of 1:1000 in blocking buffer overnight at 4°C on a rocking platform. Beta Tubulin was detected using Beta Tubulin mouse monoclonal antibody (Product # MA5-16308, lower), followed by a Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 31430) at a dilution of 1:20000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078).

Synaptophysin Antibody in Flow Cytometry (Flow) — Flow cytometry analysis of Synaptophysin was done on PC12 cells. The cells were fixed, permeabilized and stained with a Synaptophysin mouse monoclonal antibody (Product # MA1-213, red histogram) or Mouse IgG isotype control (Product # 31903, blue histogram) at a dilution of 1:500. Black histogram represents negative control unstained cell population. After incubation of the primary antibody on ice for an hour, the cells were stained with a Goat anti-Mouse IgG (H+L) Secondary Antibody, Dylight 680 conjugate (Product # 35519) at a dilution of 1:50 for at least 30 minutes on ice. A representative 10,000 cells were acquired for each sample.

Product Details

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

1:200-1:500

Immunohistochemistry (Frozen) (IHC (F))

1:100-1:500

Immunohistochemistry (Paraffin) (IHC (P))

1:100-1:500

Immunohistochemistry (PFA fixed) (IHC (PFA))

Assay Dependent

Western Blot (WB)

1:1000

Product Specifications

Species Reactivity

Human, Mouse, Rat

Host / Isotype

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

SY38

Immunogen

Synaptophysin from presynaptic vesicles from bovine brain

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A

Storage buffer

PBS

Contains

0.05% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_2723681

Product Specific Information

MA1-213 has been successfully used in western blot, imunofluorescence, immunohistochemistry, flow cytometry, immunoprecipitation and ELISA.

MA1-213 detects Synaptophysin in human, bovine, mouse and rat samples.

Target Information

Synaptophysin is a calcium-binding and integral membrane glycoprotein present in presynaptic vesicles in almost all neurons. Synaptophysin has four transmembrane domains and it forms a complex with dynamin at high calcium concentrations suggesting an involvement in synaptic vesicle endocytosis. It is also involved in the regulation of short-term and long-term synaptic plasticity. Synaptophysin is currently the most widely used marker for nerve terminals and for differentiating neuroendocrine tumors. Mutations in the gene can result in mental retardation, X-linked 96.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.