AXL Protein, Mouse (HEK293, His-Fc)
Based on 1 Customer Validation
The AXL protein has predicted roles in various cellular functions, binds to phosphatidylinositol 3-kinase and phosphatidylserine, and serves as a viral receptor. It negatively regulates apoptosis and positively regulates protein kinase B signaling. AXL Protein, Mouse (HEK293, His-Fc) is the recombinant mouse-derived AXL protein, expressed by HEK293 , with C-hFc, C-His labeled tag.
- Species: Mouse
- Source: HEK293
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Storage:Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.
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Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Technical Parameters
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Product Properties
- Documentation
- Help & FAQs
Biological Activity
Description
The AXL protein has predicted roles in various cellular functions, binds to phosphatidylinositol 3-kinase and phosphatidylserine, and serves as a viral receptor. It negatively regulates apoptosis and positively regulates protein kinase B signaling. AXL Protein, Mouse (HEK293, His-Fc) is the recombinant mouse-derived AXL protein, expressed by HEK293 , with C-hFc, C-His labeled tag.
Background
AXL protein is predicted to play a crucial role in various cellular functions, including phosphatidylinositol 3-kinase binding activity, phosphatidylserine binding activity, and virus receptor activity. Involved in negative regulation of apoptotic processes and positive regulation of protein kinase B signaling, AXL acts upstream of diverse processes such as animal organ development, myeloid cell homeostasis, and negative regulation of tumor necrosis factor production. Predicted to be located in cellular components like the actin cytoskeleton, cell surface, and host cell surface, AXL is expected to be an integral component of the plasma membrane and part of a receptor complex. With broad expression observed in various structures such as the alimentary system, brain, genitourinary system, respiratory system, and visual system, AXL likely plays a crucial role in numerous physiological processes across different tissues.
Verified Bioactivity
1.This product does not contain protein kinase domain.
2.Immobilized AXL at 2 μg/mL (100 μL/well) can bind Biotinylated GAS6 protein. The ED50 for this effect is 9.284 ng/mL, corresponding to a specific activity is 1.08×10^5 Unit/mg.
MCE Validation Data
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Purity - SDS-PAGE
Purity - SDS-PAGE
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Bioactivity - ELISA
Bioactivity - ELISA
Assay Procedure
Materials
Biotin
Pre-chilled sterile water
Wash buffer: 1X PBST
Blocking buffer: 1% BSA in 1X PBST
TMB substrate solution: Mix substrate solution A and B at a 1:1 ratio immediately before use
Stop solution: 2 M H2SO4
AXL Protein, Mouse (HEK293, His-Fc) (HY-P74406)
GAS6 Protein, Human (HEK293, C-His) (HY-P70780A)
Procedure
Biotinylation
1. Dissolution of Biotin Reagent: Remove Biotin from -10 to -30°C and immediately dissolve 1 tube of Biotin in 180 μL of cold purified water. Gently pipette to mix and obtain a 10 mM Biotin solution.
2. Calculate the volume of EZ-LinkTM Sulfo-NHS-LC-LC-Biotin, No-WeighTM Format to be added.
3. Incubation: Incubate at room temperature in the dark for 45 minutes.
4. Quantification of Biotinylated Human GAS6: Use the BCA assay to quantify the protein concentration.
ELISA Detection
1. Coating: Dilute Fibronectin Protein to 10 μg/mL in 1X PBST, add 100 μL/well, and incubate overnight at 2-8°C.
2. Washing: Wash the plate with wash buffer (260 μL/well) , once, and pat dry.
3. Blocking: Add 100 μL/well of blocking buffer, incubate at 25°C with shaking at 450 rpm for 4 hours.
4. Dilution of Biotinylated Human GAS6: Prepare dilutions at concentrations of 50, 12.5, 6.25, 3.125, 1.5625, 0.78125, 0.390625, 0.1953125, 0.097656, 0.0488, 0.0244, 0.0122, 0 μg/mL.
5. Add Biotinylated Human GAS6: Add 100 μL/well, incubate at 25°C with shaking at 450 rpm for 2 hours.
6. Washing: Wash the plate with wash buffer (260 μL/well) , five times, with each wash lasting 1 minute, then pat dry.
7. Add secondary antibody: Add 100 μL/well, incubate at 25°C with shaking at 450 rpm for 1 hour.
8. Washing: Wash the plate with wash buffer (260 μL/well) , three times, with each wash lasting 1 minute, then pat dry.
9. Add TMB Substrate Solution: Add 100 μL/well, incubate at room temperature in the dark for 15 minutes.
10. Stop Reaction: Add 100 μL of 2 M H2SO4 per well.
11. Read Plate: Measure absorbance at 450 nm.
12. Using GraphPad Prism software, plot a curve with the OD value as the vertical axis and the Biotinylated Human GAS6 protein concentration as the horizontal axis, and calculate the ED50 value.
Technical Parameters
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Species Mouse
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Source HEK293
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Tag C-hFc;C-His
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Accession
NP_033491.2 (H20-P443)
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Molecular Construction
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N-term
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AXL (H20-P443)
Accession # NP_033491.2 -
hFc-His
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C-term
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Protein Length
Extracellular Domain
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Synonyms
AXL; Tyrosine-Protein Kinase Receptor UFO; AXL Receptor Tyrosine Kinase; AXL Oncogene; UFO; Receptor Protein-Tyrosine Kinase; JTK11; AXL Transforming Sequence/Gene; Tyro7; AXL3; ARK
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AA Sequence
HKDTQTEAGSPFVGNPGNITGARGLTGTLRCELQVQGEPPEVVWLRDGQILELADNTQTQVPLGEDWQDEWKVVSQLRISALQLSDAGEYQCMVHLEGRTFVSQPGFVGLEGLPYFLEEPEDKAVPANTPFNLSCQAQGPPEPVTLLWLQDAVPLAPVTGHSSQHSLQTPGLNKTSSFSCEAHNAKGVTTSRTATITVLPQRPHHLHVVSRQPTELEVAWTPGLSGIYPLTHCNLQAVLSDDGVGIWLGKSDPPEDPLTLQVSVPPHQLRLEKLLPHTPYHIRISCSSSQGPSPWTHWLPVETTEGVPLGPPENVSAMRNGSQVLVRWQEPRVPLQGTLLGYRLAYRGQDTPEVLMDIGLTREVTLELRGDRPVANLTVSVTAYTSAGDGPWSLPVPLEPWRPGQGQPLHHLVSEPPPRAFSWP
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Molecular Weight
Approximately 100-110 kDa, based on SDS-PAGE under reducing conditions, due to the glycosylation.
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Glycosylation
Yes
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Purity
≥ 90%, as determined by reducing SDS-PAGE.
Product Properties
Lyophilized powder.
Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4.
<1 EU/μg, determined by LAL method.
It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH2O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).
Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.
Room temperature in continental US; may vary elsewhere.
Documentation
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Data Sheet (238 KB)
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SDS (251 KB)
- English - EN (251 KB)
- Français - FR (251 KB)
- Deutsch - DE (251 KB)
- Norwegian - NO (251 KB)
- Español - ES (251 KB)
- Swedish - SV (251 KB)
- Italian - IT (251 KB)
- Korean - KR (251 KB)
- Portuguese - PT (251 KB)
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Handling Instructions (2659 KB)
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
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