人間 Beta-arrestin-2 (ARRB2) ELISA Kit

商品名:	Human Beta-arrestin-2 (ARRB2) ELISA Kit
製品コード:	HUEB2090
エイリアス:	Beta-arrestin-2, Arrestin beta-2, ARRB2, ARB2, ARR2
Uniprot:	P32121
反応性:	Human
範囲:	0.312-20 ng/mL
検出方法:	Sandwich
サイズ:	96 Assay
保管所:	Please see kit components below for exact storage details
ノート:	For research use only

UniProt Protein Function:	ARRB2: a member of the arrestin/beta-arrestin protein family. These proteins participate in agonist-mediated desensitization of G-protein-coupled receptors. Acts as a cofactor in the beta-adrenergic receptor kinase (BARK) mediated desensitization of beta-adrenergic receptors. Expressed at high levels in the CNS and peripheral blood leukocytes.
UniProt Protein Details:	Protein type:Adaptor/scaffold Chromosomal Location of Human Ortholog: 17p13 Cellular Component: postsynaptic membrane; endocytic vesicle; basolateral plasma membrane; postsynaptic density; cytoplasm; plasma membrane; dendritic spine; coated pit; cytoplasmic vesicle; cytosol; nucleus Molecular Function:angiotensin receptor binding; protein domain specific binding; protein kinase B binding; type 2A serotonin receptor binding; follicle stimulating hormone receptor binding; alpha-1B adrenergic receptor binding; alpha-1A adrenergic receptor binding; protein binding; enzyme binding; G-protein-coupled receptor binding; ubiquitin protein ligase binding; protein complex binding; mitogen-activated protein kinase binding; type 1 angiotensin receptor binding; protein complex scaffold; D1 dopamine receptor binding; receptor binding Biological Process: transcription from RNA polymerase II promoter; proteasomal ubiquitin-dependent protein catabolic process; follicle-stimulating hormone signaling pathway; positive regulation of receptor internalization; protein ubiquitination; negative regulation of caspase activity; adult walking behavior; protein transport; receptor internalization; transforming growth factor beta receptor signaling pathway; negative regulation of interleukin-6 production; detection of temperature stimulus involved in sensory perception of pain; arrestin mediated desensitization of G-protein coupled receptor protein signaling pathway; platelet activation; negative regulation of interleukin-12 production; Notch signaling pathway; negative regulation of natural killer cell mediated cytotoxicity; positive regulation of synaptic transmission, dopaminergic; negative regulation of tumor necrosis factor production; negative regulation of interleukin-1 beta production; positive regulation of peptidyl-serine phosphorylation; positive regulation of protein kinase B signaling cascade; positive regulation of peptidyl-tyrosine phosphorylation; G-protein coupled receptor internalization; inhibition of NF-kappaB transcription factor; positive regulation of protein ubiquitination; negative regulation of toll-like receptor signaling pathway; negative regulation of protein ubiquitination; brain development; positive regulation of calcium ion transport; blood coagulation
NCBI Summary:	Members of arrestin/beta-arrestin protein family are thought to participate in agonist-mediated desensitization of G-protein-coupled receptors and cause specific dampening of cellular responses to stimuli such as hormones, neurotransmitters, or sensory signals. Arrestin beta 2, like arrestin beta 1, was shown to inhibit beta-adrenergic receptor function in vitro. It is expressed at high levels in the central nervous system and may play a role in the regulation of synaptic receptors. Besides the brain, a cDNA for arrestin beta 2 was isolated from thyroid gland, and thus it may also be involved in hormone-specific desensitization of TSH receptors. Multiple alternatively spliced transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Mar 2012]
UniProt Code:	P32121
NCBI GenInfo Identifier:	20141230
NCBI Gene ID:	409
NCBI Accession:	P32121.2
UniProt Related Accession:	P32121
Molecular Weight:	~ 52kDa
NCBI Full Name:	Beta-arrestin-2
NCBI Synonym Full Names:	arrestin beta 2
NCBI Official Symbol:	ARRB2
NCBI Official Synonym Symbols:	ARB2; ARR2; BARR2
NCBI Protein Information:	beta-arrestin-2
UniProt Protein Name:	Beta-arrestin-2
UniProt Synonym Protein Names:	Arrestin beta-2
Protein Family:	Beta-arrestin
UniProt Gene Name:	ARRB2
UniProt Entry Name:	ARRB2_HUMAN

成分	額	保管所
ELISA Microplate (Dismountable)	8×12 strips	-20°C
Lyophilized Standard	2	-20°C
Sample Diluent	20ml	-20°C
Assay Diluent A	10mL	-20°C
Assay Diluent B	10mL	-20°C
Detection Reagent A	120µL	-20°C
Detection Reagent B	120µL	-20°C
Wash Buffer	30mL	4°C
Substrate	10mL	4°C
Stop Solution	10mL	4°C
Plate Sealer	5	-

その他の必要な材料と設備：

Microplate reader with 450 nm wavelength filter
Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
Incubator
Deionized or distilled water
Absorbent paper
Buffer resevoir

*ノート: プロトコルは、各バッチ/ロットに固有です。正しい手順については、キットに含まれているプロトコルに従ってください。

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.

ステップ	プロトコル
1.	Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.
2.	Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.
3.	Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4.	Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.
5.	Repeat the wash process for five times as conducted in step 3.
6.	Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.
7.	Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8.	Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.
9.	After experiment, store all reagents according to the specified storage temperature respectively until their expiry.

ELISAアッセイを実施する場合、可能な限り最良の結果を達成するためにサンプルを準備することが重要です。以下に、さまざまなサンプルタイプのサンプルを準備するための手順のリストを示します。

サンプルタイプ:	プロトコル:
血清:	If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.
プラズマ:	Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
尿および脳脊髄液:	Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
細胞培養上清:	Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
細胞溶解物:	Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
組織ホモジネート:	The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
組織溶解物:	Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
母乳:	Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

ELISA	Antibodies
Human ARR beta2 (Arrestin Beta 2) CLIA Kit (HUES00358)	Anti-ARRB2 Antibody (CAB1171)
Human ARR beta2 (Arrestin Beta 2) ELISA Kit (HUES01702)	Anti-ARRB2 Antibody (CAB13278)
	Anti-ARRB2 Antibody (CAB16764)
	ARRB1 Antibody (PACO07865)
	AOAH Antibody, Biotin conjugated (PACO49401)