The Mouse Nodal ELISA Kit is a reliable and efficient tool for the quantitative measurement of Nodal levels in mouse serum, plasma, and cell culture supernatants. This kit guarantees accurate and consistent results, making it suitable for various research applications.Nodal is a pivotal protein responsible for various developmental processes, including embryogenesis and stem cell differentiation.
Dysregulation of Nodal signaling has been associated with developmental defects and diseases such as cancer and infertility, highlighting its importance as a biomarker for studying these conditions and exploring therapeutic interventions.Get your hands on the Mouse Nodal ELISA Kit today and unlock valuable insights into the role of Nodal in mouse physiology and pathology.
Product Name:
Mouse Nodal (Nodal) ELISA Kit
SKU:
MOEB1045
Size:
96T
Target:
Mouse Nodal (Nodal)
Synonyms:
Nodal, Nodal
Assay Type:
Sandwich
Detection Method:
ELISA
Reactivity:
Mouse
Detection Range:
31.2-2000pg/mL
Sensitivity:
15.64 pg/mL
Intra CV:
6.4%
Inter CV:
7.1%
Linearity:
Sample
1:2
1:4
1:8
1:16
Serum(N=5)
104-113%
95-104%
107-116%
91-101%
EDTA Plasma(N=5)
95-107%
99-109%
84-94%
105-115%
Heparin Plasma(N=5)
102-112%
106-115%
108-118%
80-88%
Recovery:
Sample Type
Average(%)
Recovery Range(%)
Serum
91
80-80
Plasma
93
80-80
Function:
Essential for mesoderm formation and axial patterning during embryonic development.
Uniprot:
P43021
Sample Type:
Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:
Natural and recombinant mouse Nodal
Sub Unit:
Homodimer; disulfide-linked.
Subcellular Location:
Secreted
Storage:
Please see kit components below for exact storage details
Note:
For research use only
UniProt Protein Function:
NODAL: Essential for mesoderm formation and axial patterning during embryonic development. Defects in NODAL are the cause of visceral heterotaxy autosomal type 5 (HTX5). A form of visceral heterotaxy, a complex disorder due to disruption of the normal left-right asymmetry of the thoracoabdominal organs. It results in an abnormal arrangement of visceral organs, and a wide variety of congenital defects. Clinical features of visceral heterotaxy autosomal type 5 include situs inversus viscerum or situs ambiguus, congenital heart defect, transposition of the great vessels ventricular septal defect, atrial septal defect, truncuscommunis, and dextrocardia. Belongs to the TGF-beta family.Protein type: Secreted; Secreted, signal peptide; Motility/polarity/chemotaxis; Cytokine; Cell development/differentiationCellular Component: extracellular region; extracellular spaceMolecular Function: cytokine activity; growth factor activity; protein binding; receptor agonist activity; transforming growth factor beta receptor bindingBiological Process: anatomical structure formation; anterior/posterior axis specification; anterior/posterior pattern formation; axial mesodermal cell fate specification; BMP signaling pathway; brain development; cell fate commitment; cell migration; cell migration involved in gastrulation; determination of left/right symmetry; digestive tract morphogenesis; embryonic cranial skeleton morphogenesis; embryonic heart tube development; embryonic pattern specification; embryonic placenta development; embryonic process involved in female pregnancy; endoderm development; floor plate morphogenesis; formation of anatomical boundary; gastrulation; gastrulation with mouth forming second; heart development; heart looping; in utero embryonic development; liver development; lung development; maternal placenta development; maternal process involved in parturition; mesendoderm development; mesoderm formation; multicellular organismal development; negative regulation of cell differentiation; negative regulation of transcription from RNA polymerase II promoter; nervous system development; neural fold formation; placenta development; polarity specification of proximal/distal axis; positive regulation of activin receptor signaling pathway; positive regulation of angiogenesis; positive regulation of caspase activity; positive regulation of cell proliferation; positive regulation of cell-cell adhesion; positive regulation of epithelial cell proliferation; positive regulation of transcription factor activity; positive regulation of transcription from RNA polymerase II promoter; regulation of apoptosis; regulation of gastrulation; regulation of MAPKKK cascade; regulation of signal transduction; repression of premature neural plate formation; stem cell maintenance; tissue morphogenesis; transforming growth factor beta receptor signaling pathway; trophectodermal cell differentiation; trophectodermal cellular morphogenesis; vasculature development
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.
