Phospho-Glycogen Synthase 1/GYS1 ( Ser 641) Rabbit mAb

Catalog No.: F2962

サイズ（液体）	価格(税別)	在庫状況
50uL	JPY 23500	国内在庫なし(納期7~10日)
100uL	JPY 35500	国内在庫なし(納期7~10日)
100uL*2	JPY 53500	お問い合わせ

代表番号: 045-509-1970\|電子メール：sales@selleck.co.jp よく尋ねられる質問

使用情報

Dilution
WB1:2000 IHC1:100-1:250

Application
WB, IHC

Source
Rabbit

Reactivity
Human, Mouse

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

Predicted MW Observed MW
84 kDa 85 kDa
^*なぜ予測分子量と実際の分子量が異なるのか？下記の原因により、実際の分子量が予測と異なる：タンパク質の翻訳後修飾（リン酸化/糖鎖付加），スプライシングバリアント，イソフォーム，相対的な電荷，ポリマー。

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
Phospho-Glycogen Synthase 1/GYS1 (Ser 641) Rabbit mAb detects endogenous levels of Glycogen Synthase 1/GYS1 only when phosphorylated at Ser 641.

Clone
E2C19

Background
Phospho-Glycogen Synthase 1/GYS1 (Ser 641) is a key enzyme in muscle glycogen biosynthesis, catalyzing the transfer of glucose from UDP-glucose to glycogen via α-1,4-glycosidic bonds. Structurally, GYS1 is a tetrameric enzyme with each subunit containing a Rossmann fold domain. It is primarily expressed in muscle but also found in the brain and other tissues. GYS1 is tightly regulated by phosphorylation, particularly at Ser641, which stabilizes an inactive conformation by interacting with an "arginine cradle" in the regulatory helices. This phosphorylation is mediated by kinases like GSK3, responding to cellular energy status and insulin signaling, ensuring glycogen synthesis occurs only when energy and glucose availability permit. Dephosphorylation or allosteric activation by glucose-6-phosphate (G6P) reactivates GYS1, linking its function to metabolic control. As a therapeutic target, GYS1 modulation is relevant in glycogen storage diseases, metabolic disorders, and conditions like Pompe disease and Lafora disease, where glycogen metabolism is disrupted.

Background

Phospho-Glycogen Synthase 1/GYS1 (Ser 641) is a key enzyme in muscle glycogen biosynthesis, catalyzing the transfer of glucose from UDP-glucose to glycogen via α-1,4-glycosidic bonds. Structurally, GYS1 is a tetrameric enzyme with each subunit containing a Rossmann fold domain. It is primarily expressed in muscle but also found in the brain and other tissues. GYS1 is tightly regulated by phosphorylation, particularly at Ser641, which stabilizes an inactive conformation by interacting with an "arginine cradle" in the regulatory helices. This phosphorylation is mediated by kinases like GSK3, responding to cellular energy status and insulin signaling, ensuring glycogen synthesis occurs only when energy and glucose availability permit. Dephosphorylation or allosteric activation by glucose-6-phosphate (G6P) reactivates GYS1, linking its function to metabolic control. As a therapeutic target, GYS1 modulation is relevant in glycogen storage diseases, metabolic disorders, and conditions like Pompe disease and Lafora disease, where glycogen metabolism is disrupted.

References
[1] McCorvie TJ, et al. Nat Struct Mol Biol. 2022 Jul;29(7):628-638. [2] Marr L, et al. Nat Commun. 2022 Jun 11;13(1):3372.

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

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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%