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Mouse Epidermal growth factor receptor (Egfr) ELISA Kit (MOEB0577)

SKU:
MOEB0577
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q01279
Range:
78-5000 pg/ml
ELISA Type:
Sandwich
Synonyms:
EGFR, ErbB-1, ErbB1, ERBB, HER1, Mena
Reactivity:
Mouse
€649

Description

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Mouse Epidermal growth factor receptor (Egfr) ELISA Kit

The Mouse Epidermal Growth Factor Receptor (EGFR) ELISA Kit is specifically designed for the accurate measurement of EGFR levels in mouse serum, plasma, and tissue lysates. This kit offers high sensitivity and specificity, providing researchers with reliable and reproducible results for a variety of research applications.EGFR is a key protein involved in cell growth, proliferation, and survival. Dysregulation of EGFR signaling has been implicated in various diseases, including cancer and inflammatory disorders.

By accurately measuring EGFR levels, researchers can gain valuable insights into disease pathogenesis and potential therapeutic targets.This ELISA kit is user-friendly and requires minimal hands-on time, making it a convenient tool for studying EGFR biology in mouse models. Whether investigating signaling pathways, drug responses, or disease mechanisms, the Mouse EGFR ELISA Kit is an essential resource for researchers looking to advance their understanding of EGFR biology.

Product Name:Mouse Epidermal growth factor receptor (Egfr) ELISA Kit
SKU:MOEB0577
Size:96T
Target:Mouse Epidermal growth factor receptor (Egfr)
Synonyms:Epidermal growth factor receptor, Egfr, 2.7.10.1
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Mouse
Detection Range:78-5000pg/ml
Sensitivity:39pg/mL
Intra CV:6.4%
Inter CV:8.1%
Linearity:
Sample1:21:41:81:16
Serum(N=5)105-117%89-99%100-113%102-112%
EDTA Plasma(N=5)99-111%96-107%92-102%84-97%
Heparin Plasma(N=5)82-94%82-91%84-94%106-115%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum9892-104
Plasma10094-106
Function:Receptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses. Known ligands include EGF, TGFA/TGF-alpha, amphiregulin, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF. Ligand binding triggers receptor homo- and/or heterodimerization and autophosphorylation on key cytoplasmic residues. The phosphorylated receptor recruits adapter proteins like GRB2 which in turn activates complex downstream signaling cascades. Activates at least 4 major downstream signaling cascades including the RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLCgamma-PKC and STATs modules. May also activate the NF-kappa-B signaling cascade. Also directly phosphorylates other proteins like RGS16, activating its GTPase activity and probably coupling the EGF receptor signaling to the G protein-coupled receptor signaling. Also phosphorylates MUC1 and increases its interaction with SRC and CTNNB1/beta-catenin. Plays a role in enhancing learning and memory performance (PubMed:20639532).
Uniprot:Q01279
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant mouse Epidermal growth factor receptor
Sub Unit:Binding of the ligand triggers homo- and/or heterodimerization of the receptor triggering its autophosphorylation. Heterodimer with ERBB2. Interacts with ERRFI1; inhibits dimerization of the kinase domain and autophosphorylation. Part of a complex with ERBB2 and either PIK3C2A or PIK3C2B. Interacts with GRB2; an adapter protein coupling the receptor to downstream signaling pathways. Interacts with GAB2; involved in signaling downstream of EGFR. Interacts with STAT3; mediates EGFR downstream signaling in cell proliferation. Interacts with RIPK1; involved in NF-kappa-B activation. Interacts (autophosphorylated) with CBL, CBLB and CBLC; involved in EGFR ubiquitination and regulation. Interacts with SOCS5; regulates EGFR degradation through ELOC- and ELOB-mediated ubiquitination and proteasomal degradation. Interacts with PRMT5; methylates EGFR and enhances interaction with PTPN6. Interacts (phosphorylated) with PTPN6; inhibits EGFR-dependent activation of MAPK/ERK. Interacts with COPG1; essential for regulation of EGF-dependent nuclear transport of EGFR by retrograde trafficking from the Golgi to the ER. Interacts with TNK2; this interaction is dependent on EGF stimulation and kinase activity of EGFR. Interacts with PCNA; positively regulates PCNA. Interacts with PELP1. Interacts with MUC1. Interacts with AP2M1. Interacts with FER. Interacts (via SH2 domains) with GRB2, NCK1 and NCK2. Interacts with EPS8; mediates EPS8 phosphorylation. Interacts with ATX2. Interacts with GAREM1. Interacts (ubiquitinated) with ANKRD13A/B/D; the interaction is direct and may regulate EGFR internalization after EGF stimulation. Interacts with GPER1; the interaction occurs in an estrogen-dependent manner. Interacts (via C-terminal cytoplasmic kinase domain) with ZPR1 (via zinc fingers). Interacts with RNF115 and RNF126. Interacts with GPRC5A (via its transmembrane domain) (PubMed:25744720). Interacts with FAM83B; positively regulates EGFR inducing its autophospharylation in absence of stimulation by EGF.
Research Area:Cancer
Subcellular Location:Cell membrane Single-pass type I membrane protein Endoplasmic reticulum membrane Single-pass type I membrane protein Golgi apparatus membrane Single-pass type I membrane protein Nucleus membrane Single-pass type I membrane protein Endosome Endosome membrane Nucleus In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER. Endocytosed upon activation by ligand. Colocalized with GPER1 in the nucleus of estrogen agonist-induced cancer-associated fibroblasts (CAF) (By similarity).
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:EGFR: a receptor tyrosine kinase. This is a receptor for epidermal growth factor (EGF) and related growth factors including TGF-alpha, amphiregulin, betacellulin, heparin-binding EGF-like growth factor, GP30, and vaccinia virus growth factor. EGFR is involved in the control of cell growth and differentiation. It is a single-pass transmembrane tyrosine kinase. Ligand binding to this receptor results in receptor dimerization, autophosphorylation (in trans), activation of various downstream signaling molecules and lysosomal degradation. It can be phosphorylated and activated by Src. Activated EGFR binds the SH2 domain of phospholipase C-gamma (PLC-gamma), activating PLC-gamma-mediated downstream signaling. Phosphorylated EGFR binds Cbl, leading to its ubiquitination and degradation. Grb2 and SHC bind to phospho-EGFR and are involved in the activation of MAP kinase signaling pathways. Phosphorylation on Ser and Thr residues is thought to represent a mechanism for attenuation of EGFR kinase activity. EGFR is overexpressed in breast, head and neck cancers, correlating with poor survival. Activating somatic mutations are seen in lung cancer, corresponding to the minority of patients with strong responses to the EGFR inhibitor Iressa (gefitinib). Mutations and amplifications are also seen in glioblastoma, and upregulation is seen in colon cancer and neoplasms. In xenografts, inhibitors synergize with cytotoxic drugs in the inhibition of many tumor types. Inhibitors include: Iressa/ZD1839, Erbitux, Tarceva, and lapatinib. Four alternatively spliced isoforms have been described.
UniProt Protein Details:

Protein type:EC 2.7.10.1; EGFR family; Kinase, protein; Membrane protein, integral; Protein kinase, TK; Protein kinase, tyrosine (receptor); TK group; Tumor suppressor

Chromosomal Location of Human Ortholog: 11 A2|11 9.41 cM

Cellular Component: apical plasma membrane; basolateral plasma membrane; cell surface; cytoplasm; early endosome membrane; endocytic vesicle; endosome; endosome membrane; focal adhesion; intracellular; lipid raft; membrane; nucleus; perinuclear region of cytoplasm; plasma membrane; receptor complex; synapse

Molecular Function:actin filament binding; cadherin binding; calmodulin binding; chromatin binding; enzyme binding; epidermal growth factor binding; epidermal growth factor receptor activity; glycoprotein binding; identical protein binding; integrin binding; kinase activity; nitric-oxide synthase regulator activity; protein binding; protein heterodimerization activity; protein kinase binding; protein phosphatase binding; protein-tyrosine kinase activity; receptor binding; signal transducer activity; transmembrane receptor activity; ubiquitin protein ligase binding

Biological Process: activation of MAPKK activity; astrocyte activation; cell morphogenesis; cell proliferation; cell surface receptor linked signal transduction; cell-cell adhesion; cerebral cortex cell migration; digestive tract morphogenesis; embryonic placenta development; epidermal growth factor receptor signaling pathway; epidermis development; G1/S-specific positive regulation of cyclin-dependent protein kinase activity; hair follicle development; learning and/or memory; negative regulation of apoptosis; negative regulation of mitotic cell cycle; negative regulation of protein catabolic process; neurite morphogenesis; ovulation cycle; peptidyl-tyrosine phosphorylation; positive regulation of bone resorption; positive regulation of cell growth; positive regulation of cell migration; positive regulation of cell proliferation; positive regulation of DNA repair; positive regulation of DNA replication; positive regulation of epithelial cell proliferation; positive regulation of fibroblast proliferation; positive regulation of inflammatory response; positive regulation of MAP kinase activity; positive regulation of nitric oxide biosynthetic process; positive regulation of phosphorylation; positive regulation of protein amino acid phosphorylation; positive regulation of protein kinase B signaling cascade; positive regulation of smooth muscle cell proliferation; positive regulation of superoxide release; positive regulation of synaptic transmission, glutamatergic; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription, DNA-dependent; positive regulation of vasoconstriction; protein amino acid autophosphorylation; receptor-mediated endocytosis; regulation of cell proliferation; regulation of JNK cascade; regulation of nitric-oxide synthase activity; regulation of peptidyl-tyrosine phosphorylation; regulation of phosphoinositide 3-kinase cascade; response to calcium ion; salivary gland morphogenesis; signal transduction; translation; wound healing

UniProt Code:Q01279
NCBI GenInfo Identifier:1352359
NCBI Gene ID:13649
NCBI Accession:Q01279.1
UniProt Related Accession:Q01279
Molecular Weight:134,853 Da
NCBI Full Name:Epidermal growth factor receptor
NCBI Synonym Full Names:epidermal growth factor receptor
NCBI Official Symbol:Egfr
NCBI Official Synonym Symbols:Wa5; wa2; Erbb; Errp; wa-2; Errb1; AI552599; 9030024J15Rik
NCBI Protein Information:epidermal growth factor receptor
UniProt Protein Name:Epidermal growth factor receptor
Protein Family:Epidermal growth factor receptor
UniProt Gene Name:Egfr
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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