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Rat Cadherin-2 (Cdh2) ELISA Kit (RTEB0998)

SKU:
RTEB0998
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q9Z1Y3
ELISA Type:
Sandwich
Synonyms:
NCAD, Cadherin-2, CD325
Reactivity:
Rat
€649
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Description

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Rat Cadherin-2 (Cdh2) ELISA Kit

The Rat Cadherin-2 (CDH2) ELISA Kit is specifically designed for the quantitative detection of CDH2 levels in rat samples, including serum, plasma, and tissue homogenates. This kit offers high sensitivity and specificity, ensuring accurate and reliable results for your research needs.Cadherin-2, also known as N-cadherin, is a cell surface glycoprotein that plays a crucial role in cell adhesion and signaling. It is involved in various biological processes, including development, tissue organization, and wound healing. Dysregulation of CDH2 expression has been linked to several diseases, such as cancer, Alzheimer's disease, and cardiovascular disorders, highlighting its importance as a potential biomarker for disease research and therapeutic development.

With easy-to-follow protocols and rapid assay procedures, the Rat Cadherin-2 (CDH2) ELISA Kit is the perfect tool for studying the role of CDH2 in physiological and pathological processes, providing valuable insights into disease mechanisms and potential treatment options. Choose this kit for robust and accurate quantification of CDH2 levels in rat samples.

Product Name:Rat Cadherin-2 (Cdh2) ELISA Kit
SKU:RTEB0998
Size:96T
Target:Rat Cadherin-2 (Cdh2)
Synonyms:Neural cadherin, N-cadherin, CD325
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Rat
Detection Range:0.312-20ng/mL
Sensitivity:0.192ng/mL
Intra CV:Provided with the Kit
Inter CV:Provided with the Kit
Linearity:Provided with the Kit
Recovery:Provided with the Kit
Function:Cadherins are calcium-dependent cell adhesion proteins. They preferentially interact with themselves in a homophilic manner in connecting cells; cadherins may thus contribute to the sorting of heterogeneous cell types. Acts as a regulator of neural stem cells quiescence by mediating anchorage of neural stem cells to ependymocytes in the adult subependymal zone: upon cleavage by MMP24, CDH2-mediated anchorage is affected, leading to modulate neural stem cell quiescence. CDH2 may be involved in neuronal recognition mechanism. In hippocampal neurons, may regulate dendritic spine density.
Uniprot:Q9Z1Y3
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant rat Cadherin-2
Sub Unit:Interacts with CDCP1 (By similarity). Interacts with PCDH8; this complex may also include TAOK2 (PubMed:17988630). The interaction with PCDH8 may lead to internalization through TAOK2/p38 MAPK pathway (PubMed:17988630). Identified in a complex containing FGFR4, NCAM1, CDH2, PLCG1, FRS2, SRC, SHC1, GAP43 and CTTN (By similarity). May interact with OBSCN (via protein kinase domain 2).
Subcellular Location:Cell membrane Single-pass type I membrane protein Colocalizes with TMEM65 at the intercalated disk in cardiomyocytes. Colocalizes with OBSCN at the intercalated disk and sarcolemma in cardiomyocytes.
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:CDH2: Cadherins are calcium dependent cell adhesion proteins. They preferentially interact with themselves in a homophilic manner in connecting cells; cadherins may thus contribute to the sorting of heterogeneous cell types. CDH2 may be involved in neuronal recognition mechanism. In hippocampal neurons, may regulate dendritic spine density. Interacts with CDCP1. Identified in a complex containing FGFR4, NCAM1, CDH2, PLCG1, FRS2, SRC, SHC1, GAP43 and CTTN. Interacts with PCDH8; this complex may also include TAOK2. The interaction with PCDH8 may lead to internalization through TAOK2/p38 MAPK pathway.
UniProt Protein Details:

Protein type:Membrane protein, integral; Cell adhesion; Motility/polarity/chemotaxis

Cellular Component: focal adhesion; protein complex; basolateral plasma membrane; T-tubule; integral to membrane; intercellular junction; catenin complex; fascia adherens; cell-cell adherens junction; adherens junction; membrane; lamellipodium; apical plasma membrane; plasma membrane; synapse

Molecular Function:protein binding; enzyme binding; gamma-catenin binding; beta-catenin binding; protein complex binding; calcium ion binding; nitric-oxide synthase binding; protein phosphatase binding; protein kinase binding; alpha-catenin binding

Biological Process: cell migration; regulation of myelination; protein heterooligomerization; regulation of axonogenesis; heterophilic cell adhesion; cell-cell adhesion; synaptogenesis; glial cell differentiation; regulation of protein localization; positive regulation of MAPKKK cascade; calcium-dependent cell-cell adhesion; regulation of Rho protein signal transduction; striated muscle cell differentiation; blood vessel morphogenesis; cell adhesion; homophilic cell adhesion

NCBI Summary:cell-cell adhesion molecule; may play a role in smooth muscle cell migration and adhesion in response to injury [RGD, Feb 2006]
UniProt Code:Q9Z1Y3
NCBI GenInfo Identifier:13431334
NCBI Gene ID:83501
NCBI Accession:Q9Z1Y3.1
UniProt Secondary Accession:Q9Z1Y3,Q9R0T5,
UniProt Related Accession:Q9Z1Y3
Molecular Weight:99,686 Da
NCBI Full Name:Cadherin-2
NCBI Synonym Full Names:cadherin 2
NCBI Official Symbol:Cdh2
NCBI Official Synonym Symbols:N-cadherin
NCBI Protein Information:cadherin-2; neural cadherin; cadherin 2 type 1 N-cadherin (neuronal); cadherin 2, type 1, N-cadherin (neuronal)
UniProt Protein Name:Cadherin-2
UniProt Synonym Protein Names:Neural cadherin; N-cadherin; CD_antigen: CD325
UniProt Gene Name:Cdh2
UniProt Entry Name:CADH2_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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