The ImmunoStar VMAT2 antiserum was quality control tested using standard immunohistochemical methods in rat brain and adrenal medulla using biotin/avidin-HRP techniques.
Specificity of the antiserum was demonstrated by soluble pre-adsorption and Western blot. Tissue staining is completely eliminated by pretreatment of the diluted antibody with an excess of rat VMAT2 peptide residues 496-515.
Western blot analysis of immunoprecipitated rat brain homogenates demonstrates a dense immunoreactive band of approximately 55 kD and a minor band of approximately 75 kD.
Photo Description: IHC image of VMAT2 staining in the rat adrenal medulla (above) and of neurons in the pontine region of the rat brain (below). The tissue was fixed with 4% formaldehyde in 0.1 M phosphate buffer, before being removed and prepared for vibratome sectioning for rat brain and frozen sectioning for adrenal gland. Sections were incubated at RT in 10% goat serum in PBS, before standard IHC procedure. Primary antibody was incubated at 1:5000 for 48 hours, goat anti-rabbit secondary was subsequently added for 1 hour after washing with PBS. Light microscopy staining was achieved with standard biotin-streptavidin/HRP procedure and DAB chromogen.
Quantity / Volume: 100 µL
State: Lyophilized Whole Serum
Species Reactivity: Rat
Availability: In Stock
Alternate Names: Monoamine transporter; Solute carrier family 18 member 2; Synaptic vesicular amine transporter; Vesicular amine transporter 2; Solute carrier family 18 A2 (vesicular monoamine transporter 2); SLC18A2; SVMT; VAT2; Svat; mnat, anti-VMAT2
Gene Symbol: Slc18a2
Entrez Gene: 25549 Rat
This product contains the preservative sodium azide. The concentration percent of the sodium azide is ≤ .09%. Although this hazardous substance is a concentration below that required for the preparation of a Material Safety Data Sheet, we created a standard MSDS for your records.
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Works Very Well
We have used the VMAT2 antibody from your company a couple of times.
From the results it seems that this antibody works very well: it labels the structures in the brain and the spinal cord where this transporter should be located and labeling is very clear.
Mengliang Zhang, DM, PhD, Associate Professor
University of Copenhagen
Department of Neuroscience and Pharmacology
The Panum Institute, bldg 33.3
I have used this antibody (#20042) to label the brains of adult Mouse. Works very well. I highly recommend. I used the antibody on mouse Striatal and substantia Nigra tissue for fluorescence microscopy. I tried different concentration and 1:2000 works the best. The staining was also done in transgenic mice that express GFP on TH promoter for a positive control. The entire process was done on free-floating sections by keeping sections on the shaker agitating and slow speed.
Mice were cardiac perfused with fresh 4% Paraformaldehyde and 15% picric acid in 0.1M PB and post fix in same fixative overnight at 4ÂºC.
-Washed brain 3 x 10min in phosphate buffered saline (PBS; pH 7.4)
-Brains were cut @ 50um on vibratome
-Clean sections with 1% sodium borohydride in PBS for 15 min @ RT, followed by 3 x 10min PBS wash.
-Quenched endogenous reaction with 10% Methanol and 3% Hydrogen peroxide in PBS for 15 min @ RT, followed by 3 x 10min PBS wash.
-Blocked with 3% bovine serum albumin (BSA), 10% Normal Donkey serum (NDS) and 0.5% Triton X in PBS 4-6 hours @ RT.
– Incubate in primary (Rb anti-VMAT2, 1:2000; Immunostar #20042) in 1% BSA, 1% NDS and 0.1% Triton-X in PBS for 60-70 hours @ 4ÂºC (This is over the weekend. For fast result, can also incubate for overnight @ RT).
Day 4 or 6
-Wash 3 x 10 min in PBS
– Incubate in secondary (Donkey Anti Rabbit IgG with Alexa 594, 1:400) for 4-6 hours @ RT
-Wash 3 x 10min in PBS
-Mount section and cover slip
Laboratory Researcher III