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Apelin-12 (Human, Rat, Mouse, Bovine) - Antibody for Immunohistochemistry

Catalog# Standard size Price Order
H-057-23 50 l $450.00

Image



Apelin-like immunoreactivity (Apelin-12 (Human, Mouse, Rat, Bovine) Antiserum, Catalog No.: H-057-23, Phoenix Pharmaceuticals) vascular endothelial cells of large conduit vessels. Photomicrographs show staining in saphenous vein (a) and coronary artery (c) with higher magnification of the respective vessels in (b) and (d).
Kleinz et al. Regul Pept. 2004 May 15;118(3):119-25. 

Immunocytochemistry Methods:
Cryostat cut tissue sections (10 Am for atrial and ventricular myocardium, lung, kidney and adrenal gland. 30 Am for saphenous vein, radial artery, left internal mammary artery and coronary artery) were left to dry overnight at room temperature. Tissue was fixed in ice-cold acetone for 10 min. Sections were incubated with 5% non-immunized swine serum in PBS for 1 h at room temperature to block nonspecific protein interaction. Tissues were incubated with rabbit-anti-apelin-12 (human/rat) antiserum at a dilution of 1:500 in PBS/T containing 1% non-immunized swine serum (1%SS PBS/T) for 72 h at 4oC. Tissue sections were then washed three times for 5 min in cold PBS/T before the incubation with swine-anti-rabbit antiserum at a dilution of 1:200 in 1%SS PBS/T for 1 h at room temperature. After repeated washing, sections were incubated with rabbit peroxidase/anti-peroxidase complex at a 1:400 dilution in 1%SS PBS/T. Further washing preceded a 3-min incubation of tissue sections with a 2,5% solution of 3,3V-diaminobenzidine in 0.05M Tris-HCl buffer containing 0.3% hydrogen peroxide. The chromogenic reaction was stopped by immersing the slides in distilled water. Sections were dehydrated using a graded alcohol series before being submerged in xylene for 1 h to clear. Sections were mounted using DePeXGurr mounting medium and then examined using a standard light field microscope (Reichert-Jung, Austria).
Kleinz et al. Regul Pept. 2004 May 15;118(3):119-25.

     

Protocol for Immunohistochemistry: 
 Tissue Sample Rat lateral hypothalamus & medial forebrain boundle
 Fixative 10% Formalin
 Embedding Paraffin
 Negative control No primary antibody
 Pretreatment Target Retrieval 25 min (Steam)
 Blocking 2% Normal Goat Serum
 Primary Antibody Rabbit Anti-Apelin-12 (Human, Bovine) Antiserum (Catalog No.:H-057-23)
 Optimal Dilution  1:3000 (1hour at RT)
 Secondary Antibody Goat anti-Rabbit IgG, Biotinylated (1:400, 30 min)
 Amplification  Streptavidin-HRP (Vector), 1:400, 30 min
 Detection system HRP
 Substrate DAB (Sigma), 3 min
 Counterstained Hematoxylin, 30 Sec

Catalog #H-057-23
Standard Size50 l
SequenceArg - Pro - Arg - Leu - Ser - His - Lys - Gly - Pro - Met - Pro - Phe
SpeciesHuman, Bovine, Rat, Mouse
HostRabbit
ReconstitutionFor best and reproducible results, reconstitute with 50l of distilled water for the equivalent of undiluted antiserum, immediately before use. Do not refreeze any unused portions.
Storage ConditionPlease store the lyophilized antibody at -20°C upon receipt. For optimal results, use the antibody immediately after reconstitution. Once reconstituted, the antibody is stable for up to three (3) days at 4°C. For longer-term storage up to three (3) months, prepare small aliquots of the reconstituted antibody and freeze at -20°C or -80°C. Repeated freeze-thaw cycles should be strictly avoided.
ContentThis vial contains 50µl of Rabbit Anti-Apelin-12 (Human, Bovine) Serum in the lyophilized form.
Recommended Dilution FactorImmunocytochemistry - 1:500. Regul Pept. 2004 May 15;118(3):119-25. 
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