Immunofluorescence analysis of GHRL was performed using 70% confluent log phase MKN-45 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Ghrelin Rabbit Polyclonal Antibody (PA1-1046) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues G(1) S S F L S P E H Q R V Q Q R K E S K K P P A K L P Q R(28) of human GHRL.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||1 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-1046 detects recombinant human ghrelin. This antibody detect both the octanoylated and non-octanoylated forms of ghrelin.
PA1-1046 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~3 kDa protein representing recombinant human ghrelin.
The PA1-1046 immunogen is a synthetic peptide corresponding to residues G(1) S S F L S P E H Q R V Q Q R K E S K K P P A K L P Q R(28) of human GHRL.
Ghrelin was originally discovered as a preprohormone that stimulates the release of growth hormone from the anterior pituitary. Recent studies have shown that ghrelin, in concert with other hormones, helps to control both appetite and energy balance.
The synthesis of ghrelin takes place predominantly in epithelial cells of the stomach, although it is also produced in smaller quantities in the placenta, kidney, pituitary and hypothalamus. Ghrelin is produced as a preprohormone that is proteolytically cleaved to yield its active form, a 28-amino acid peptide. The further octanoylation modification of the ghrelin peptide is being implicated in the regulation of the hormone.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.