target post-translational modification
unmodified
conjugate
unconjugated
antibody form
affinity purified immunoglobulin
clone
polyclonal
species reactivity
rat, rabbit
concentration
(Please refer to the Certificate of Analysis for the lot-specific concentration.)
technique(s)
western blot: suitable
UniProt accession no.
Quality Level
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General description
FUNCTION: SwissProt: P30872 # Receptor for somatostatin with higher affinity for somatostatin-14 than -28. This receptor is coupled via pertussis toxin sensitive G proteins to inhibition of adenylyl cyclase. In addition it stimulates phosphotyrosine phosphatase and Na(+)/H(+) exchanger via pertussis toxin insensitive G proteins.
SIZE: 391 amino acids; 42686 Da
SUBUNIT: Interacts with SKB1.
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Fetal kidney, fetal liver, and adult pancreas, brain, lung, jejunum and stomach.
SIMILARITY: SwissProt: P30872 ## Belongs to the G-protein coupled receptor 1 family.
SIZE: 391 amino acids; 42686 Da
SUBUNIT: Interacts with SKB1.
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Fetal kidney, fetal liver, and adult pancreas, brain, lung, jejunum and stomach.
SIMILARITY: SwissProt: P30872 ## Belongs to the G-protein coupled receptor 1 family.
Purified peptide from amino acids 26-40 of rat SSTR1 (Accession number P28646). The immunogen sequence is identical in mouse and 13/15 identical in human.
Application
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).
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).
Biochem/physiol Actions
Recognizes Somatostatin Receptor Type 1 (SSTR1).
Physical form
Affinity purified immunoglobulin. Lyophilized from phosphate buffered saline, pH 7.4, containing 1% BSA, and 0.05% sodium azide as a preservative. Reconstitute with 200 μL of sterile deionized water. Centrifuge antibody preparation before use (10,000 xg for 5 min).
Preparation Note
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.
Other Notes
Included free of charge with the antibody is 40 μg of control antigen (lyophilized powder). The stock solution of the antigen can be made up using 100 μL of sterile deionized 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.
Western blot: 1:200 using ECL on rat brain lysate. The antibody recognizes a single band around 55-60kDa.
Dilutions should be made using a carrier protein such as BSA (1-3%)
Optimal working dilutions must be determined by the end user.
Dilutions should be made using a carrier protein such as BSA (1-3%)
Optimal working dilutions must be determined by the end user.
Disclaimer
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.
存储类别
10 - Combustible liquids
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