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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| R | WB | Rb | Affinity Purified | Polyclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | AB9018-50UL |
| Brand Family | Chemicon® |
| Trade Name |
|
| Description | Anti-Muscarinic Acetylcholine Receptor m3 Antibody |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Control |
|
| Presentation | 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). |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This Anti-Muscarinic Acetylcholine Receptor m3 Antibody is validated for use in WB for the detection of Muscarinic Acetylcholine Receptor m3. |
| Key Applications |
|
| Application Notes | 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). |
| Biological Information | |
|---|---|
| Immunogen | Synthetic peptide from the 3rd intracellular loop of rat m3 (Accession P08483). The immunogen sequence is identical in human, mouse, bovine and porcine. |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rabbit |
| Specificity | Muscarinic acetylcholine receptor m3. |
| Species Reactivity |
|
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
|
| Entrez Gene Summary | The muscarinic cholinergic receptors belong to a larger family of G protein-coupled receptors. The functional diversity of these receptors is defined by the binding of acetylcholine and includes cellular responses such as adenylate cyclase inhibition, phosphoinositide degeneration, and potassium channel mediation. Muscarinic receptors influence many effects of acetylcholine in the central and peripheral nervous system. The muscarinic cholinergic receptor 3 controls smooth muscle contraction and its stimulation causes secretion of glandular tissue. |
| Gene Symbol |
|
| UniProt Number |
|
| UniProt Summary | FUNCTION: SwissProt: P20309 # The muscarinic acetylcholine receptor mediates various cellular responses, including inhibition of adenylate cyclase, breakdown of phosphoinositides and modulation of potassium channels through the action of G proteins. Primary transducing effect is Pi turnover. SIZE: 590 amino acids; 66128 Da SUBCELLULAR LOCATION: Basolateral cell membrane; Multi-pass membrane protein. SIMILARITY: SwissProt: P20309 ## Belongs to the G-protein coupled receptor 1 family. |
| Product Usage Statements | |
|---|---|
| Usage Statement |
|
| Storage and Shipping Information | |
|---|---|
| Storage Conditions | 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. |
| Packaging Information | |
|---|---|
| Material Size | 50 µL |