Invitrogen

Synaptophysin Monoclonal Antibody (SY38)

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This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

1 Published Figure

2 References

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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.

Synaptophysin Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across tissues tested owing to their inherent genetic constitution. Relative expression of Synaptophysin was observed in Mouse Brain in comparison to Mouse Heart and Mouse Liver using Anti-Synaptophysin Monoclonal Antibody (SY38) (Product # MA1-213) in Western Blot. {RE}

Synaptophysin Antibody in Immunocytochemistry, Immunohistochemistry (ICC/IF, IHC) — FIGURE 7 Impaired rod maturation in PDE6B patient retinal organoids. (A) Representative immunostaining pictures of the presynaptic marker synaptophysin and rhodopsin (RHO) in retinal organoids. Bar: 25 mum. (B) Amplified presynaptic formation of rod photoreceptors indicated with arrows in (A) . Bar: 25 mum. (C) Colocalization of RHO and synaptophysin. (D) Representative immunostaining pictures of the cilium marker ARL13B and RHO in retinal organoids at D230. Bar: 25 mum. Boxes were amplified images. White arrows indicated definite signal of ARL13B represent cilia. Bar: 6.25 mum. (E) Statistic analysis of cilia number in retinal organoids at D230. Statistical results are the mean values +- SD of 3 organoids (3 images each) (D230) of two independent differentiations from the control and patient. Two-tailed t -test, *** P < 0.001.

Product Details

MA1-213

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000-1:1,500	-
Immunohistochemistry (IHC)	-	View 1 publication 1 publication
Immunohistochemistry (Paraffin) (IHC (P))	1:100-1:500	-
Immunohistochemistry (PFA fixed) (IHC (PFA))	Assay-dependent	-
Immunohistochemistry (Frozen) (IHC (F))	1:100-1:500	-
Immunocytochemistry (ICC/IF)	-	View 1 publication 1 publication
Flow Cytometry (Flow)	1:200-1:500	-

Product Specifications
Species Reactivity	Bovine, Human, Mouse, Rat
Published species	Not Applicable
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.