Tyrosine-protein kinase receptor UFO is a protein that in human is encoded by the AXL gene. The gene was initially designated as UFO, in allusion to the unidentified function of this protein. However, in the years since its discovery, research into AXL's expression profile and mechanism has made it an increasingly attractive target, especially for cancer therapeutics. In recent years, AXL has emerged as a key facilitator of immune escape and drug-resistance by cancer cells, leading to aggressive and metastatic cancers. AXL is a cell surface receptor tyrosine kinase, part of the TAM family of kinases including TYRO3 and MERTK.
Background References
1. Tanaka M et al. Therapeutic Targeting of the Gas6/Axl Signaling Pathway in Cancer. Int J Mol Sci. 2021 Sep
2. Adam-Artigues A et al. AXL - a new player in resistance to HER2 blockade. Cancer Treat Rev. 2023 Dec
Western blot analysis of Axl on different lysates with Rabbit anti-Axl antibody (HA722740) at 1/2,000 dilution.
Lane 1: NCI-H1299 cell lysate Lane 2: Jurkat cell lysate (negative) Lane 3: HeLa cell lysate Lane 4: HUVEC cell lysate Lane 5: COS-1 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 98 kDa Observed band size: 140 kDa
Exposure time: 59 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722740) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Relative expression (RE)
Immunocytochemistry analysis of NCI-H1299 (positive) and Jurkat (negative) labeling Axl with Rabbit anti-Axl antibody (HA722740) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Axl antibody (HA722740) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
☑ Relative expression (RE)
Flow cytometric analysis of Jurkat (left, negative) and NCI-H1299 (right, positive) cells labeling Axl.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA722740, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"