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Rat Gastrin/cholecystokinin type B receptor (Cckbr) ELISA Kit (RTEB0607)

SKU:
RTEB0607
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P30553
ELISA Type:
Sandwich
Reactivity:
Rat
$779
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Description

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Rat Gastrin/cholecystokinin type B receptor (Cckbr) ELISA Kit

The Rat Gastrin Cholecystokinin Type B Receptor (CCKBR) ELISA Kit is a powerful tool for studying the expression levels of CCKBR in rat samples. This kit offers high sensitivity and specificity, allowing for accurate detection of CCKBR in serum, plasma, and cell culture supernatants. CCKBR is a key receptor involved in the regulation of various physiological processes, including digestion, appetite regulation, and stress responses. Dysregulation of CCKBR signaling has been implicated in conditions such as obesity, gastrointestinal disorders, and anxiety disorders, making it a valuable target for research and therapeutic development.

By utilizing the Rat CCKBR ELISA Kit, researchers can gain valuable insights into the role of CCKBR in health and disease, paving the way for the development of novel treatments and interventions. With its reliable performance and ease of use, this kit is an essential tool for advancing our understanding of CCKBR biology.

Product Name:Rat Gastrin/cholecystokinin type B receptor (Cckbr) ELISA Kit
SKU:RTEB0607
Size:96T
Target:Rat Gastrin/cholecystokinin type B receptor (Cckbr)
Synonyms:Cholecystokinin-2 receptor, CCK2-R, CCK-B receptor
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Rat
Detection Range:31.2-2000pg/mL
Sensitivity:15pg/mL
Intra CV:Provided with the Kit
Inter CV:Provided with the Kit
Linearity:Provided with the Kit
Recovery:Provided with the Kit
Function:Receptor for gastrin and cholecystokinin. The CKK-B receptors occur throughout the central nervous system where they modulate anxiety, analgesia, arousal, and neuroleptic activity. This receptor mediates its action by association with G proteins that activate a phosphatidylinositol-calcium second messenger system.
Uniprot:P30553
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant rat Gastrin/cholecystokinin type B receptor
Subcellular Location:Cell membrane Multi-pass membrane protein
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:CCKBR: Receptor for gastrin and cholecystokinin. The CKK-B receptors occur throughout the central nervous system where they modulate anxiety, analgesia, arousal, and neuroleptic activity. This receptor mediates its action by association with G proteins that activate a phosphatidylinositol-calcium second messenger system. Belongs to the G-protein coupled receptor 1 family. 3 isoforms of the human protein are produced by alternative splicing.Protein type: Membrane protein, multi-pass; Receptor, GPCR; Membrane protein, integral; GPCR, family 1Cellular Component: membrane; cytoplasm; plasma membrane; integral to membrane; nucleusMolecular Function: type B gastrin/cholecystokinin receptor binding; gastrin receptor activityBiological Process: gland development; gastric acid secretion; apoptosis; positive regulation of synaptic transmission, GABAergic; behavioral defense response; positive regulation of synaptic transmission, glutamatergic; positive regulation of synaptic transmission; response to insulin stimulus; histamine secretion; G-protein coupled receptor protein signaling pathway; elevation of cytosolic calcium ion concentration; gut development; positive regulation of cell proliferation; digestion; regulation of sensory perception of pain
UniProt Protein Details:
NCBI Summary:Cholecystokinin was one of the first gastrointestinal peptides discovered in the mammalian brain. Cholecystokinin receptors are members of the G protein-coupled receptor superfamily, stimulating phosphatidylinositol turnover and intracellular calcium mobilization. This gene encodes a cholecystokinin receptor of the B subtype. These receptors occur throughout the central nervous system where they modulate anxiety, analgesia, arousal, and neuroleptic activity. [provided by RefSeq, Jul 2008]
UniProt Code:P30553
NCBI GenInfo Identifier:6978617
NCBI Gene ID:25706
NCBI Accession:NP_037297.1
UniProt Secondary Accession:P30553
UniProt Related Accession:P30553
Molecular Weight:48,957 Da
NCBI Full Name:gastrin/cholecystokinin type B receptor
NCBI Synonym Full Names:cholecystokinin B receptor
NCBI Official Symbol:Cckbr
NCBI Official Synonym Symbols:Cck2r; Cholrec
NCBI Protein Information:gastrin/cholecystokinin type B receptor; CCK-BR; CCK2-R; CCK2 receptor; CCK-B receptor; CCK(B) receptor; cholecystokinin-2 receptor
UniProt Protein Name:Gastrin/cholecystokinin type B receptor
UniProt Synonym Protein Names:Cholecystokinin-2 receptor
Protein Family:Gastrin/cholecystokinin type B receptor
UniProt Gene Name:Cckbr
UniProt Entry Name:GASR_RAT
Component Quantity (96 Assays) Storage
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 -

Other materials and equipment required:

  • 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

*Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

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.

Step
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.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type Protocol
Serum 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.
Plasma 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.
Urine & Cerebrospinal Fluid 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.
Cell culture supernatant 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.
Cell lysates 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.
Tissue homogenates 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.
Tissue lysates 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.
Breast Milk 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.

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