Ghrelin Antibody [F2E5]

Catalog No.: F3727

Application: Reactivity: Mouse, Rat, Human

Immunohistochemical analysis of formalin fixed paraffin embedded mouse stomach tissue with F3727 at 1:8000 dilution.

当該製品は品切れ状态で、メールアドレスをご教示いただければ、お客様に返信いたします。

代表番号: 045-509-1970|電子メール：sales@selleck.co.jp

使用情報

Dilution
IHC1:8000

Application
IHC

Source
Rabbit Monoclonal Antibody

Reactivity
Mouse, Rat, Human

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

ポジティブコントロール	Mouse stomach tissue; Human stomach tissue; Rat stomach tissue
ネガティブコントロール	Rat liver tissue; Mouse liver tissue; Human liver tissue

プロトコール

IHC
Experimental Protocol： Deparaffinization/Rehydration 1. Deparaffinize/hydrate sections: 2. Incubate sections in three washes of xylene for 5 min each. 3. Incubate sections in two washes of 100% ethanol for 10 min each. 4. Incubate sections in two washes of 95% ethanol for 10 min each. 5. Wash sections two times in dH2O for 5 min each. 6.Antigen retrieval: For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; continue with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min. Staining 1. Wash sections in dH2O three times for 5 min each. 2. Incubate sections in 3% hydrogen peroxide for 10 min. 3. Wash sections in dH2O two times for 5 min each. 4. Wash sections in wash buffer for 5 min. 5. Block each section with 100–400 µl of blocking solution for 1 hr at room temperature. 6. Remove blocking solution and add 100–400 µl primary antibody diluent in to each section. Incubate overnight at 4°C. 7. Remove antibody solution and wash sections with wash buffer three times for 5 min each. 8. Cover section with 1–3 drops HRPas needed. Incubate in a humidified chamber for 30 min at room temperature. 9. Wash sections three times with wash buffer for 5 min each. 10. Add DAB Chromogen Concentrate to DAB Diluent and mix well before use. 11. Apply 100–400 µl DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity. 12. Immerse slides in dH2O. 13. If desired, counterstain sections with hematoxylin. 14. Wash sections in dH2O two times for 5 min each. 15. Dehydrate sections: Incubate sections in 95% ethanol two times for 10 sec each; Repeat in 100% ethanol, incubating sections two times for 10 sec each; Repeat in xylene, incubating sections two times for 10 sec each. 16. Mount sections with coverslips and mounting medium.

IHC

Experimental Protocol：

Deparaffinization/Rehydration

1. Deparaffinize/hydrate sections:

2. Incubate sections in three washes of xylene for 5 min each.

3. Incubate sections in two washes of 100% ethanol for 10 min each.

4. Incubate sections in two washes of 95% ethanol for 10 min each.

5. Wash sections two times in dH2O for 5 min each.

6.Antigen retrieval: For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; continue with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.

Staining

1. Wash sections in dH2O three times for 5 min each.

2. Incubate sections in 3% hydrogen peroxide for 10 min.

3. Wash sections in dH2O two times for 5 min each.

4. Wash sections in wash buffer for 5 min.

5. Block each section with 100–400 µl of blocking solution for 1 hr at room temperature.

6. Remove blocking solution and add 100–400 µl primary antibody diluent in to each section. Incubate overnight at 4°C.

7. Remove antibody solution and wash sections with wash buffer three times for 5 min each.

8. Cover section with 1–3 drops HRPas needed. Incubate in a humidified chamber for 30 min at room temperature.

9. Wash sections three times with wash buffer for 5 min each.

10. Add DAB Chromogen Concentrate to DAB Diluent and mix well before use.

11. Apply 100–400 µl DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity.

12. Immerse slides in dH2O.

13. If desired, counterstain sections with hematoxylin.

14. Wash sections in dH2O two times for 5 min each.

15. Dehydrate sections: Incubate sections in 95% ethanol two times for 10 sec each; Repeat in 100% ethanol, incubating sections two times for 10 sec each; Repeat in xylene, incubating sections two times for 10 sec each.

16. Mount sections with coverslips and mounting medium.

Datasheet & SDS

生物学的記述

Specificity
Ghrelin Antibody [F2E5] detects endogenous levels of total Ghrelin protein.

タンパク質の局在
細胞外環境

Uniprot ID
Q9UBU3

Clone
F2E5

Synonym(s)
MTLRP; UNQ524/PRO1066; GHRL; Appetite‑regulating hormone; Growth hormone secretagogue; Growth hormone‑releasing peptide; Motilin‑related peptide; Protein M46

Background
Ghrelin is an acylated peptide hormone synthesized predominantly in gastric enteroendocrine cells, with additional expression in the pancreas, lung, and hypothalamic neurons, where it acts as a key peripheral and central regulator of energy homeostasis, growth hormone release, and reward related feeding behavior. The mature ligand binds to the growth hormone secretagogue receptor (GHSR/GHS R1a), a class A G protein–coupled receptor highly expressed in the hypothalamic arcuate nucleus, pituitary, and midbrain dopamine circuits, engaging multiple G protein families including Gαq/11, Gαi/o, and Gα12/13, which activate phospholipase C–inositol trisphosphate–calcium signaling, inhibit adenylate cyclase, and recruit Rho A–dependent pathways, respectively, while β arrestins additionally mediate GHSR internalization and MAPK activation. In the arcuate nucleus, ghrelin GHSR signaling stimulates neuropeptide Y and agouti related peptide–expressing orexigenic neurons, increases AMP activated protein kinase activity, and disinhibits dopaminergic reward pathways, thereby promoting hunger, anticipatory feeding, and preference for calorie dense foods, whereas in the pituitary the same receptor ligand pair drives calcium dependent growth hormone secretion, linking ghrelin to somatotropic axis control and metabolic adaptation. Ghrelin also modulates gastric motility, acid secretion, and pancreatic islet function. Ghrelin-mediated Gαi signalling attenuates glucose-stimulated insulin release, thereby coordinating energy intake, storage, and fuel utilisation under fasted states. In contrast, its circulating levels normally decline after meals to disinhibit satiety pathways governed by leptin and insulin. In obesity, ghrelin suppression can blunt meal-induced inhibition of appetite and impair metabolic flexibility. Conversely, in cachexia and chronic illness, reduced ghrelin tone may exacerbate negative energy balance and muscle wasting, ultimately leading to dysregulated ghrelin-GHSR signalling across both hyperphagic and catabolic metabolic states.

Background

Ghrelin is an acylated peptide hormone synthesized predominantly in gastric enteroendocrine cells, with additional expression in the pancreas, lung, and hypothalamic neurons, where it acts as a key peripheral and central regulator of energy homeostasis, growth hormone release, and reward related feeding behavior. The mature ligand binds to the growth hormone secretagogue receptor (GHSR/GHS R1a), a class A G protein–coupled receptor highly expressed in the hypothalamic arcuate nucleus, pituitary, and midbrain dopamine circuits, engaging multiple G protein families including Gαq/11, Gαi/o, and Gα12/13, which activate phospholipase C–inositol trisphosphate–calcium signaling, inhibit adenylate cyclase, and recruit Rho A–dependent pathways, respectively, while β arrestins additionally mediate GHSR internalization and MAPK activation. In the arcuate nucleus, ghrelin GHSR signaling stimulates neuropeptide Y and agouti related peptide–expressing orexigenic neurons, increases AMP activated protein kinase activity, and disinhibits dopaminergic reward pathways, thereby promoting hunger, anticipatory feeding, and preference for calorie dense foods, whereas in the pituitary the same receptor ligand pair drives calcium dependent growth hormone secretion, linking ghrelin to somatotropic axis control and metabolic adaptation. Ghrelin also modulates gastric motility, acid secretion, and pancreatic islet function. Ghrelin-mediated Gαi signalling attenuates glucose-stimulated insulin release, thereby coordinating energy intake, storage, and fuel utilisation under fasted states. In contrast, its circulating levels normally decline after meals to disinhibit satiety pathways governed by leptin and insulin. In obesity, ghrelin suppression can blunt meal-induced inhibition of appetite and impair metabolic flexibility. Conversely, in cachexia and chronic illness, reduced ghrelin tone may exacerbate negative energy balance and muscle wasting, ultimately leading to dysregulated ghrelin-GHSR signalling across both hyperphagic and catabolic metabolic states.

References
[1] Kirchner H, et al. Handb Exp Pharmacol. 2012;(209):161-84. [2] Lv Y, et al. Biosci Rep. 2018 Sep 25;38(5):BSR20181061.

PVDFメンブレン孔径参考表

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

技術サポート

ストックの作り方、阻害剤の保管方法、細胞実験や動物実験の際に注意すべき点など、製品を取扱う時に問い合わせが多かった質問に対しては取扱説明書でお答えしています。

Handling Instructions

他に質問がある場合は、お気軽にお問い合わせください。

* 必須

* 大学・企業名 大学・企業名を記入してください

* 名前 名前を記入してください

* 電子メール 電子メール・アドレスを記入してください 有効なメールアドレスを入力してください

電話番号

* 内容 お問い合わせ内容をご入力ください

Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%