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Mouse Tyrosine-protein kinase receptor UFO (Axl) ELISA Kit (MOEB2106)

SKU:
MOEB2106
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q00993
ELISA Type:
Sandwich
Reactivity:
Mouse
€649
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Description

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Mouse Tyrosine-protein kinase receptor UFO (Axl) ELISA Kit

The Mouse Tyrosine Protein Kinase Receptor UFO (AXL) ELISA Kit is a valuable tool for the accurate measurement of AXL levels in mouse serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit ensures precise and dependable results, making it an excellent choice for various research applications.AXL, also known as UFO, is a tyrosine protein kinase receptor that plays a critical role in cell growth, survival, and migration. It has been implicated in cancer progression, immune response regulation, and tissue regeneration, making it a key target for studying various diseases and potential therapeutic interventions.

This ELISA kit provides researchers with a reliable method for quantifying AXL levels in mouse samples, allowing for in-depth investigations into the role of this protein in different biological processes and disease pathways. Get accurate and reproducible results with the Mouse Tyrosine Protein Kinase Receptor UFO (AXL) ELISA Kit.

Product Name:Mouse Tyrosine-protein kinase receptor UFO (Axl) ELISA Kit
SKU:MOEB2106
Size:96T
Target:Mouse Tyrosine-protein kinase receptor UFO (Axl)
Synonyms:Adhesion-related kinase, Ark, Ufo
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Mouse
Detection Range:31.2-2000pg/mL
Sensitivity:15.62pg/mL
Intra CV:6.3%
Inter CV:7.8%
Linearity:
Sample1:21:41:81:16
Serum(N=5)90-99%85-97%99-110%99-109%
EDTA Plasma(N=5)92-104%98-110%111-121%109-118%
Heparin Plasma(N=5)101-111%106-118%86-96%103-116%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum9589-101
Plasma9791-103
Function:Receptor tyrosine kinase that transduces signals from the extracellular matrix into the cytoplasm by binding growth factor GAS6 and which is thus regulating many physiological processes including cell survival, cell proliferation, migration and differentiation. Ligand binding at the cell surface induces dimerization and autophosphorylation of AXL. Following activation by ligand, ALX binds and induces tyrosine phosphorylation of PI3-kinase subunits PIK3R1, PIK3R2 and PIK3R3; but also GRB2, PLCG1, LCK and PTPN11. Other downstream substrate candidates for AXL are CBL, NCK2, SOCS1 and TNS2. Recruitment of GRB2 and phosphatidylinositol 3 kinase regulatory subunits by AXL leads to the downstream activation of the AKT kinase. GAS6/AXL signaling plays a role in various processes such as endothelial cell survival during acidification by preventing apoptosis, optimal cytokine signaling during human natural killer cell development, hepatic regeneration, gonadotropin-releasing hormone neuron survival and migration, platelet activation, or regulation of thrombotic responses. Plays also an important role in inhibition of Toll-like receptors (TLRs)-mediated innate immune response.
Uniprot:Q00993
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant mouse Tyrosine-protein kinase receptor UFO
Sub Unit:Heterodimer and heterotetramer with ligand GAS6 (By similarity). Interacts with CBL, GRB2, LCK, NCK2, PIK3R1, PIK3R2, PIK3R3, PLCG1, SOCS1 and TNS2. Part of a complex including AXL, TNK2 and GRB2, in which GRB2 promotes AXL recruitment by TNK2.
Research Area:Cancer
Subcellular Location:Cell membrane Single-pass type I membrane protein
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:AXL: a receptor tyrosine kinase that may function as a signal transducer between specific cell types of mesodermal origin. Interacts with SKP1. Overexpression in tissue culture causes oncogenic transformation. Overexpressed in several cancers including thyroid, ovarian, gastric, ER+ breast cancer and acute myeloid leukemia, where it is associated with poor prognosis. 2 isoforms of the human protein are produced by alternative splicing.Protein type: Membrane protein, integral; Protein kinase, TK; Protein kinase, tyrosine (receptor); EC 2.7.10.1; Oncoprotein; Kinase, protein; TK group; Axl familyCellular Component: extracellular space; cell surface; membrane; cell; integral to membrane; plasma membrane; intracellularMolecular Function: transferase activity; phosphatidylserine binding; protein heterodimerization activity; protein-tyrosine kinase activity; transferase activity, transferring phosphorus-containing groups; nucleotide binding; phosphoinositide 3-kinase binding; kinase activity; transmembrane receptor protein tyrosine kinase activity; ATP binding; myosin heavy chain binding; protein kinase activityBiological Process: peptidyl-tyrosine phosphorylation; neuron migration; protein amino acid phosphorylation; enzyme linked receptor protein signaling pathway; positive regulation of natural killer cell differentiation; erythrocyte homeostasis; ovulation cycle; natural killer cell differentiation; negative regulation of interferon-gamma production; protein kinase B signaling cascade; negative regulation of neuron apoptosis; inflammatory response; cell differentiation; secretion by cell; forebrain cell migration; negative regulation of lymphocyte activation; platelet activation; immune system process; vagina development; negative regulation of tumor necrosis factor production; phagocytosis; positive regulation of protein kinase B signaling cascade; cellular response to extracellular stimulus; innate immune response; spermatogenesis; blood vessel remodeling; positive regulation of cytokine and chemokine mediated signaling pathway; phosphorylation; apoptotic cell clearance; negative regulation of apoptosis
UniProt Protein Details:
NCBI Summary:
UniProt Code:Q00993
NCBI GenInfo Identifier:342187104
NCBI Gene ID:26362
NCBI Accession:Q00993.2
UniProt Secondary Accession:Q00993,Q80YQ3
UniProt Related Accession:Q00993
Molecular Weight:98,191 Da
NCBI Full Name:Tyrosine-protein kinase receptor UFO
NCBI Synonym Full Names:AXL receptor tyrosine kinase
NCBI Official Symbol:Axl
NCBI Official Synonym Symbols:Ark; Ufo; Tyro7; AI323647
NCBI Protein Information:tyrosine-protein kinase receptor UFO; ufo oncogene homolog; adhesion-related kinase
UniProt Protein Name:Tyrosine-protein kinase receptor UFO
UniProt Synonym Protein Names:Adhesion-related kinase
Protein Family:Tyrosine-protein kinase receptor
UniProt Gene Name:Axl
UniProt Entry Name:UFO_MOUSE
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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