Human cAMP-dependent protein kinase catalytic subunit beta (PRKACB) ELISA Kit (HUEB2083)
- SKU:
- HUEB2083
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P22694
- Range:
- 78-5000 pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- PRKACB, cAMP-dependent protein kinase catalytic subunit beta
- Reactivity:
- Human
Description
Human cAMP-dependent protein kinase catalytic subunit beta (PRKACB) ELISA Kit
The Human cAMP-dependent Protein Kinase Catalytic Subunit Beta (PRKACB) ELISA Kit is a highly reliable and sensitive tool for detecting PRKACB levels in human samples such as serum, plasma, and cell culture supernatants. This kit offers accurate and reproducible results, making it ideal for various research applications.PRKACB is an essential protein kinase that plays a key role in regulating cellular processes such as cell growth, metabolism, and gene expression.
Dysregulation of PRKACB has been implicated in various diseases, including cancer, diabetes, and neurodegenerative disorders, making it a valuable biomarker for studying these conditions and potential therapeutic interventions.With its high sensitivity and specificity, the Human PRKACB ELISA Kit from AssayGenie is a valuable tool for researchers seeking to explore the role of PRKACB in health and disease.
| Product Name: | Human cAMP-dependent protein kinase catalytic subunit beta (PRKACB) ELISA Kit |
| SKU: | HUEB2083 |
| Size: | 96T |
| Target: | Human cAMP-dependent protein kinase catalytic subunit beta (PRKACB) |
| Synonyms: | PKA C-beta |
| Assay Type: | Sandwich |
| Detection Method: | ELISA |
| Reactivity: | Human |
| Detection Range: | 78-5000pg/mL |
| Sensitivity: | 20pg/mL |
| Intra CV: | 5.9% | ||||||||||||||||||||
| Inter CV: | 10.9% | ||||||||||||||||||||
| Linearity: |
| ||||||||||||||||||||
| Recovery: |
| ||||||||||||||||||||
| Function: | Mediates cAMP-dependent signaling triggered by receptor binding to GPCRs. PKA activation regulates diverse cellular processes such as cell proliferation, the cell cycle, differentiation and regulation of microtubule dynamics, chromatin condensation and decondensation, nuclear envelope disassembly and reassembly, as well as regulation of intracellular transport mechanisms and ion flux. Regulates the abundance of compartmentalized pools of its regulatory subunits through phosphorylation of PJA2 which binds and ubiquitinates these subunits, leading to their subsequent proteolysis. |
| Uniprot: | P22694 |
| Sample Type: | Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids |
| Specificity: | Natural and recombinant human cAMP-dependent protein kinase catalytic subunit beta |
| Sub Unit: | A number of inactive tetrameric holoenzymes are produced by the combination of homo- or heterodimers of the different regulatory subunits associated with two catalytic subunits. cAMP causes the dissociation of the inactive holoenzyme into a dimer of regulatory subunits bound to four cAMP and two free monomeric catalytic subunits. The cAMP-dependent protein kinase catalytic subunit binds PJA2. |
| Research Area: | Cardiovascular |
| Subcellular Location: | Cytoplasm Cell membrane Membrane Lipid-anchor Nucleus Translocates into the nucleus (monomeric catalytic subunit). The inactive holoenzyme is found in the cytoplasm. |
| Storage: | Please see kit components below for exact storage details |
| Note: | For research use only |
| UniProt Protein Function: | PKACB: Mediates cAMP-dependent signaling triggered by receptor binding to GPCRs. PKA activation regulates diverse cellular processes such as cell proliferation, the cell cycle, differentiation and regulation of microtubule dynamics, chromatin condensation and decondensation, nuclear envelope disassembly and reassembly, as well as regulation of intracellular transport mechanisms and ion flux. Regulates the abundance of compartmentalized pools of its regulatory subunits through phosphorylation of PJA2 which binds and ubiquitinates these subunits, leading to their subsequent proteolysis. A number of inactive tetrameric holoenzymes are produced by the combination of homo- or heterodimers of the different regulatory subunits associated with two catalytic subunits. cAMP causes the dissociation of the inactive holoenzyme into a dimer of regulatory subunits bound to four cAMP and two free monomeric catalytic subunits. The cAMP-dependent protein kinase catalytic subunit binds PJA2. Isoform 1 is most abundant in the brain, with low level expression in kidney. Isoform 2 is predominantly expressed in thymus, spleen and kidney. Isoform 3 and isoform 4 are only expressed in the brain. Activated by cAMP. Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. cAMP subfamily. 9 isoforms of the human protein are produced by alternative splicing. |
| UniProt Protein Details: | Protein type:Protein kinase, Ser/Thr (non-receptor); Kinase, protein; Protein kinase, AGC; EC 2.7.11.11; AGC group; PKA family Chromosomal Location of Human Ortholog: 1p31.1 Cellular Component: nucleoplasm; centrosome; perinuclear region of cytoplasm; plasma membrane; cytosol; cAMP-dependent protein kinase complex Molecular Function:protein binding; cAMP-dependent protein kinase activity; ubiquitin protein ligase binding; magnesium ion binding; ATP binding Biological Process: epidermal growth factor receptor signaling pathway; fibroblast growth factor receptor signaling pathway; nerve growth factor receptor signaling pathway; water transport; activation of protein kinase A; glucose metabolic process; pathogenesis; negative regulation of meiotic cell cycle; signal transduction; protein amino acid phosphorylation; gluconeogenesis; G-protein signaling, coupled to cAMP nucleotide second messenger; synaptic transmission; phospholipase C activation; triacylglycerol catabolic process; carbohydrate metabolic process; neural tube closure; energy reserve metabolic process; renal water homeostasis; innate immune response; blood coagulation; transmembrane transport; regulation of insulin secretion |
| NCBI Summary: | The protein encoded by this gene is a member of the serine/threonine protein kinase family. The encoded protein is a catalytic subunit of cAMP (cyclic AMP)-dependent protein kinase, which mediates signalling though cAMP. cAMP signaling is important to a number of processes, including cell proliferaton and differentiation. Multiple alternatively spliced transcript variants encoding distinct isoforms have been observed. [provided by RefSeq, Jul 2014] |
| UniProt Code: | P22694 |
| NCBI GenInfo Identifier: | 125210 |
| NCBI Gene ID: | 5567 |
| NCBI Accession: | P22694.2 |
| UniProt Secondary Accession: | P22694,Q14VH1, Q59GC0, Q5BNE9, Q5BNF0, Q5BNF1, Q5BNF2 Q5BNF3, Q5CZ92, B1APG4, B4DKB0, B4E2Q1, |
| UniProt Related Accession: | P22694 |
| Molecular Weight: | 37,110 Da |
| NCBI Full Name: | cAMP-dependent protein kinase catalytic subunit beta |
| NCBI Synonym Full Names: | protein kinase, cAMP-dependent, catalytic, beta |
| NCBI Official Symbol: | PRKACB |
| NCBI Official Synonym Symbols: | PKACB; PKA C-beta |
| NCBI Protein Information: | cAMP-dependent protein kinase catalytic subunit beta; protein kinase A catalytic subunit beta; cAMP-dependent protein kinase catalytic beta subunit isoform 4ab |
| UniProt Protein Name: | cAMP-dependent protein kinase catalytic subunit beta |
| Protein Family: | cAMP-dependent protein kinase |
| UniProt Gene Name: | PRKACB |
| UniProt Entry Name: | KAPCB_HUMAN |
| 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. |
Related Products
Bovine cAMP-dependent protein kinase catalytic subunit beta (PRKACB) ELISA Kit (BOEB0939)
Porcine cAMP-dependent protein kinase catalytic subunit beta (PRKACB) ELISA Kit (PREB0526)
Rat cAMP-dependent protein kinase catalytic subunit beta (Prkacb) ELISA Kit (RTEB1310)
Mouse cAMP-dependent protein kinase catalytic subunit beta (Prkacb) ELISA Kit (MOEB1834)
Human cAMP-dependent protein kinase catalytic subunit alpha (PRKACA) ELISA Kit (HUEB2037)
Human PRKACB / PKA beta (catalytic subunit) ELISA Kit
Bovine cAMP-dependent protein kinase catalytic subunit alpha (PRKACA) ELISA Kit (BOEB0912)
Mouse cAMP-dependent protein kinase catalytic subunit alpha (Prkaca) ELISA Kit (MOEB1779)
