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Apelin-36 (Human) - Antibody for Immunohistochemistry

Catalog# Standard size Price Order
H-057-15 50 µl $450.00

Image


Rat hypothalamus was stained by Anti-Apelin-36 (Human) Serum (H-057-15)
Brailoiu et al. Neurosci Lett. 2002 Jul 26;327(3):193-7.

Protocol for Immunohistochemistry: 
 Tissue Sample Rat Hypothalamus
 Fixative 4% paraformaldehyde/ 0.2% picric acid in PBS
 Embedding Paraffin
 Negative control No primary antibody and pre-absorption of the antibody with the peptide apelin-36 (1µg/ml)
 Pretreatment No
 Blocking 2% Normal Goat Serum
 Primary Antibody Rabbit Anti-Apelin-36 (Human) Serum (Catalog No.: H-057-15)
 Optimal Dilution 1:3000, overnight at 4ºC
 Secondary Antibody Goat Anti-Rabbit IgG, Biotinylated (1:50), 30 min
 Amplification  Streptavidin-HRP (Vector), 1:400, 30 min
 Detection system HRP
 Substrate DAB (Sigma), 3 min

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 stained using rabbit-anti-apelin-36 (human) antiserum at a dilution of 1:200 in PBS/T for 24 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.

Fig. 5. Apelin-like immunoreactivity in vascular endothelial cells of large conduit vessels. Photomicrograph shows staining in saphenous vein.

 

Fig. 3. Photomicrograph demonstrating apelin-like immunoreactivity (apelin-LI) in the human heart. Apelin-LI is expressed in vascular endothelial cells of right atria.

 
Kleinz et al. Regul Pept. 2004 May 15;118(3):119-25.

 

Protocol for Immunohistochemistry: 
 Tissue Sample Human placenta tissue
 Fixative 10% Formalin
 Embedding Paraffin
 Negative control No primary antibody
 Pretreatment Intact
 Blocking 2% Normal Goat Serum
 Primary Antibody Rabbit Anti-Apelin-36 (Human) Serum (Catalog No.: H-057-15)
 Optimal Dilution 1:200 (1hour at RT)
 Secondary Antibody Goat anti-Rabbit IgG, Biotinylated (1:400)
 Amplification  Streptavidin-HRP (Vector), 1:400, 30 min
 Detection system HRP
 Substrate DAB (Sigma)
 Counterstained Hematoxylin
Catalog #H-057-15
Standard Size50 µl
SequenceLeu - Val - Gln - Pro - Arg - Gly - Ser - Arg - Asn - Gly - Pro - Gly - Pro - Trp - Gln - Gly - Gly - Arg - Arg - Lys - Phe - Arg - Arg - Gln - Arg - Pro - Arg - Leu - Ser - His - Lys - Gly - Pro - Met - Pro - Phe
SpeciesHuman
HostRabbit
ReconstitutionFor best and reproducible results, reconstitute with 50µl 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-36 (human) serum in the lyophilized form.
Recommended Dilution FactorImmunohistochemistry - 1:400. Regul Pept. 2007 Aug 16;142(3):131-7.
Immunocytochemistry - 1:200. Regul Pept. 2004 May 15;118(3):119-25.
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