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Rat Syntenin-1 (Sdcbp) ELISA Kit (RTEB1030)

SKU:
RTEB1030
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q9JI92
ELISA Type:
Sandwich
Reactivity:
Rat
€649

Description

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Rat Syntenin-1 (Sdcbp) ELISA Kit

The Rat Syntenin-1 (SDCBP) ELISA Kit is a powerful tool designed for the accurate detection of syntenin-1 levels in rat samples such as serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring precise and reliable results that are essential for various research applications. Syntenin-1 is a key protein involved in cell adhesion, migration, and signaling pathways. It plays a crucial role in various cellular processes, making it a valuable biomarker for studying diseases such as cancer, inflammatory disorders, and neurodegenerative conditions.

The Rat Syntenin-1 (SDCBP) ELISA Kit provides researchers with a reliable means to measure syntenin-1 levels, paving the way for further advancements in understanding disease mechanisms and developing potential therapeutic interventions. To learn more about the Rat Syntenin-1 (SDCBP) ELISA Kit, visit the link: www.assaygenie.com/rat-syntenin-1-sdcbp-elisa-kit/

Product Name:Rat Syntenin-1 (Sdcbp) ELISA Kit
SKU:RTEB1030
Size:96T
Target:Rat Syntenin-1 (Sdcbp)
Synonyms:Syndecan-binding protein 1
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Rat
Detection Range:0.156-10ng/mL
Sensitivity:0.065ng/mL
Intra CV:4.3%
Inter CV:7.5%
Linearity:
Sample1:21:41:81:16
Serum(N=5)101-111%94-103%102-113%82-92%
EDTA Plasma(N=5)98-106%110-120%105-117%102-112%
Heparin Plasma(N=5)105-117%105-117%108-118%92-101%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum9387-99
Plasma9589-101
Function:Multifunctional adapter protein involved in diverse array of functions including trafficking of transmembrane proteins, neuro and immunomodulation, exosome biogenesis, and tumorigenesis. Positively regulates TGFB1-mediated SMAD2/3 activation and TGFB1-induced epithelial-to-mesenchymal transition (EMT) and cell migration in various cell types. May increase TGFB1 signaling by enhancing cell-surface expression of TGFR1 by preventing the interaction between TGFR1 and CAV1 and subsequent CAV1-dependent internalization and degradation of TGFR1. In concert with SDC1/4 and PDCD6IP, regulates exosome biogenesis. Regulates migration, growth, proliferation, and cell cycle progression in a variety of cancer types. In adherens junctions may function to couple syndecans to cytoskeletal proteins or signaling components. Seems to couple transcription factor SOX4 to the IL-5 receptor (IL5RA). May also play a role in vesicular trafficking. Seems to be required for the targeting of TGFA to the cell surface in the early secretory pathway.
Uniprot:Q9JI92
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant rat Syntenin-1
Sub Unit:Monomer and homodimer. Interacts with neurofascin and SDCBP2 (PubMed:11152476). Interacts with PTPRJ (PubMed:11434923). Interacts with SDC1, SDC2, SDC3, SDC4, NRXN2, EPHA7, EPHB1, NF2 isoform 1, TGFA and IL5RA (By similarity). Interacts with PDCD6IP. Forms a complex with PDCD6IP and SDC2. Interacts (via C-terminus) with TGFBR1 (By similarity). Binds to FZD7; this interaction is increased by inositol trisphosphate (IP3).
Subcellular Location:Cell junction Focal adhesion Cell junction Adherens junction Cell membrane Peripheral membrane protein Endoplasmic reticulum membrane Peripheral membrane protein Nucleus Melanosome Cytoplasm Cytosol Cytoplasm Cytoskeleton Secreted Exosome Membrane raft Mainly membrane-associated. Localized to adherens junctions, focal adhesions and endoplasmic reticulum. Colocalized with actin stress fibers. Also found in the nucleus. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. Associated to the plasma membrane in the presence of FZD7 and phosphatidylinositol 4,5-bisphosphate (PIP2) (By similarity).
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:syntenin: Seems to function as an adapter protein. In adherens junctions may function to couple syndecans to cytoskeletal proteins or signaling components. Seems to couple transcription factor SOX4 to the IL-5 receptor (IL5RA). May also play a role in vesicular trafficking. Seems to be required for the targeting of TGFA to the cell surface in the early secretory pathway. 3 isoforms of the human protein are produced by alternative splicing.Protein type: Cytokine; Adaptor/scaffold; Motility/polarity/chemotaxis; Cell adhesion; Membrane protein, integral; VesicleCellular Component: extracellular space; cytoplasm; plasma membrane; melanosome; interleukin-5 receptor complex; intracellular; vesicleMolecular Function: syndecan binding; protein C-terminus binding; identical protein binding; protein homodimerization activity; ephrin receptor binding; protein N-terminus binding; interleukin-5 receptor binding; protein binding; frizzled binding; protein heterodimerization activity; growth factor binding; neurexin binding; cell adhesion molecule binding; glycoprotein bindingBiological Process: Ras protein signal transduction; positive regulation of phosphorylation
UniProt Protein Details:
NCBI Summary:a neurofascin-binding protein [RGD, Feb 2006]
UniProt Code:Q9JI92
NCBI GenInfo Identifier:14010891
NCBI Gene ID:83841
NCBI Accession:NP_114192.1
UniProt Secondary Accession:Q9JI92
UniProt Related Accession:Q9JI92
Molecular Weight:32,423 Da
NCBI Full Name:syntenin-1
NCBI Synonym Full Names:syndecan binding protein
NCBI Official Symbol:Sdcbp
NCBI Official Synonym Symbols:mda-9; TACIP18
NCBI Protein Information:syntenin-1
UniProt Protein Name:Syntenin-1
UniProt Synonym Protein Names:Syndecan-binding protein 1
Protein Family:Syntenin
UniProt Gene Name:Sdcbp
UniProt Entry Name:SDCB1_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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