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Merck
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Key Documents

AB9018

Sigma-Aldrich

Anti-Muscarinic Acetylcholine Receptor m3 Antibody

Chemicon®, from rabbit

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About This Item

UNSPSC 코드:
12352203
eCl@ss:
32160702
NACRES:
NA.41

생물학적 소스

rabbit

Quality Level

항체 형태

affinity purified immunoglobulin

항체 생산 유형

primary antibodies

클론

polyclonal

정제법

affinity chromatography

종 반응성

rat

제조업체/상표

Chemicon®

기술

western blot: suitable

NCBI 수납 번호

UniProt 수납 번호

배송 상태

wet ice

타겟 번역 후 변형

unmodified

유전자 정보

특이성

Muscarinic acetylcholine receptor m3.

면역원

Synthetic peptide from the 3rd intracellular loop of rat m3 (Accession P08483). The immunogen sequence is identical in human, mouse, bovine and porcine.

애플리케이션

Research Category
Neuroscience
Research Sub Category
Neurotransmitters & Receptors
This Anti-Muscarinic Acetylcholine Receptor m3 Antibody is validated for use in WB for the detection of Muscarinic Acetylcholine Receptor m3.
Western blot: 1:2000 using ECL on rat brain lysate.

Dilutions should be made using a carrier protein such as BSA (1-3%)

Optimal working dilutions must be determined by the end user.

SUGGESTED WESTERN BLOT PROTOCOL

1. Mix the samples (organ membranes: 50 μg/lane; transfected cells: 500,000 cells/lane) with sample-buffer X 2, and heat 10 min at 70°C.

2. 5-50 μL applied to Minigel lane (0.75-1.5 mm width) and run at standard conditions. (60 mA for 2 1.5 mm Minigel gels, 1.4 h). It is suggested that you run 5-15% acrylamide (37.5:1 acrylamide:bisacrysmide) minigel (1.5 mm width) at 30 mA/gel ~1-1.5 hours.

3. Transfer in semi-dry system under standard conditions (3 h 100 mA for two minigel gels)

4. Stain the transferred bands with Chemicon BLOT-FastStain (Catalog Number 2076).

5. Destain with deionized water.

6. Block with 5% non-fat milk (Marvel or Carnation) in PBS, and 0.025 % sodium azide, overnight at 2-8°C. The non-fat milk should be dissolved freshly, centrifuged 10,000 rpm for 10 min, and filtered through glass filter (Gelman Acrodisc).

7. Incubation with first antibody 2 h at room temperature or overnight at 4°C in blocking solution. The antibody preparation should be centrifuged before use (10,000 g 5 min.). Optimal working dilutions and incubation time will need to be determined by the end user.

8. Wash 4 x 10 min. with PBS-0.1% tween 20. From this stage, azide should be omitted.

9. Incubation with the secondary antibody (HRP-conjugated goat anti-rabbit antibody, for example Chemicon Catalog Number AP132P, diluted appropriately) 1 h at room temperature.

10. Wash 4 x 10 min. with PBS-0.1% tween 20.

11. Perform ECL with commercial kits (Chemilucent, Chemicon Catalog Number 2600).

물리적 형태

Affinity purified immunoglobulin. Lyophilized from phosphate buffered saline, pH 7.4, containing 1% BSA, and 0.05% sodium azide as a preservative. Reconstitute with 50 μL of sterile deionized water. Centrifuge antibody preparation before use (10,000 xg for 5 min).

저장 및 안정성

Maintain lyophilized material at -20°C for up to 12 months after date of receipt. After reconstitution maintain at -20°C in undiluted aliquots for up to 6 months. Avoid repeated freeze/thaw cycles.

분석 메모

Control
Included free of charge with the antibody is 40 μg of control antigen. The stock solution of the antigen can be made up using 100 μL of sterile distilled water. For negative control, preincubate 1 μg of peptide with 1 μg of antibody for one hour at room temperature. Optimal concentrations must be determined by the end user.

기타 정보

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

법적 정보

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

면책조항

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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유해 및 위험 성명서

예방조치 성명서

Hazard Classifications

Aquatic Chronic 3

Storage Class Code

11 - Combustible Solids

WGK

WGK 3


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