Phospho-AXL (Tyr702) Antibody - #AF8523

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製品: Phospho-AXL (Tyr702) Antibody
カタログ: AF8523
タンパク質の説明: Rabbit polyclonal antibody to Phospho-AXL (Tyr702)
アプリケーション: WB IHC IF/ICC
反応性: Human, Mouse, Rat
予測: Pig, Zebrafish, Bovine, Horse, Sheep, Rabbit, Dog, Xenopus
分子量: 97.4kDa; 98kD(Calculated).
ユニプロット: P30530
RRID: AB_2840577

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製品説明

ソース:
Rabbit
アプリケーション:
WB 1:1000-3000, IF/ICC 1:100-1:500, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

反応性:
Human,Mouse,Rat
予測:
Pig(100%), Zebrafish(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Xenopus(100%)
クローナリティ:
Polyclonal
特異性:
Phospho-AXL (Tyr702) Antibody detects endogenous levels of AXL only when phosphorylated at Tyr702.
RRID:
AB_2840577
引用形式: Affinity Biosciences Cat# AF8523, RRID:AB_2840577.
コンジュゲート:
Unconjugated.
精製:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
保存:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
別名:

折りたたみ／展開

Adhesion related kinase; AI323647; Ark; Axl; AXL oncogene; AXL receptor tyrosine kinase; AXL transforming gene; AXL transforming sequence/gene; EC 2.7.10.1; JTK11; Oncogene AXL; Tyro7; Tyrosine protein kinase receptor UFO; Tyrosine-protein kinase receptor UFO; UFO; UFO_HUMAN;

免疫原

免疫原:
Uniprot:

P30530(UFO_HUMAN) >>Visit HPA database.

遺伝子(ID):
AXL(558)
発現特異性:
P30530 UFO_HUMAN:

Highly expressed in metastatic colon tumors. Expressed in primary colon tumors. Weakly expressed in normal colon tissue.

タンパク質配列:
MAWRCPRMGRVPLAWCLALCGWACMAPRGTQAEESPFVGNPGNITGARGLTGTLRCQLQVQGEPPEVHWLRDGQILELADSTQTQVPLGEDEQDDWIVVSQLRITSLQLSDTGQYQCLVFLGHQTFVSQPGYVGLEGLPYFLEEPEDRTVAANTPFNLSCQAQGPPEPVDLLWLQDAVPLATAPGHGPQRSLHVPGLNKTSSFSCEAHNAKGVTTSRTATITVLPQQPRNLHLVSRQPTELEVAWTPGLSGIYPLTHCTLQAVLSDDGMGIQAGEPDPPEEPLTSQASVPPHQLRLGSLHPHTPYHIRVACTSSQGPSSWTHWLPVETPEGVPLGPPENISATRNGSQAFVHWQEPRAPLQGTLLGYRLAYQGQDTPEVLMDIGLRQEVTLELQGDGSVSNLTVCVAAYTAAGDGPWSLPVPLEAWRPGQAQPVHQLVKEPSTPAFSWPWWYVLLGAVVAAACVLILALFLVHRRKKETRYGEVFEPTVERGELVVRYRVRKSYSRRTTEATLNSLGISEELKEKLRDVMVDRHKVALGKTLGEGEFGAVMEGQLNQDDSILKVAVKTMKIAICTRSELEDFLSEAVCMKEFDHPNVMRLIGVCFQGSERESFPAPVVILPFMKHGDLHSFLLYSRLGDQPVYLPTQMLVKFMADIASGMEYLSTKRFIHRDLAARNCMLNENMSVCVADFGLSKKIYNGDYYRQGRIAKMPVKWIAIESLADRVYTSKSDVWSFGVTMWEIATRGQTPYPGVENSEIYDYLRQGNRLKQPADCLDGLYALMSRCWELNPQDRPSFTELREDLENTLKALPPAQEPDEILYVNMDEGGGYPEPPGAAGGADPPTQPDPKDSCSCLTAAEVHPAGRYVLCPSTTPSPAQPADRGSPAAPGQEDGA

種類予測

種類予測:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Xenopus
100
Zebrafish
100
Rabbit
100
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P30530 基板として

Site PTM Type Enzyme
S100 Phosphorylation
T200 Phosphorylation
S201 Phosphorylation
S202 Phosphorylation
S204 Phosphorylation
T303 Phosphorylation
Y481 Phosphorylation
S503 Phosphorylation
S515 Phosphorylation
K523 Ubiquitination
K525 Ubiquitination
K535 Ubiquitination
K540 Ubiquitination
K563 Ubiquitination
K570 Ubiquitination
S612 Phosphorylation
Y634 Phosphorylation
Y643 Phosphorylation
K666 Ubiquitination
S694 Phosphorylation
K696 Ubiquitination
Y698 Phosphorylation
Y702 Phosphorylation
Y703 Phosphorylation
K710 Ubiquitination
K714 Ubiquitination
R724 Methylation
Y726 Phosphorylation
Y759 Phosphorylation
Y772 Phosphorylation
Y779 Phosphorylation P30530 (AXL) , P00533 (EGFR)
Y814 Phosphorylation
Y821 Phosphorylation P30530 (AXL)
S853 Phosphorylation
Y866 Phosphorylation P30530 (AXL)
T873 Phosphorylation
S875 Phosphorylation
S884 Phosphorylation

PTMs - P30530 酵素として

Substrate Site Source
P30530-1 (AXL) Y779 Uniprot
P30530-1 (AXL) Y821 Uniprot
P30530 (AXL) Y866 Uniprot

研究背景

機能:

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.

(Microbial infection) Acts as a receptor for lassa virus and lymphocytic choriomeningitis virus, possibly through GAS6 binding to phosphatidyl-serine at the surface of virion envelope.

(Microbial infection) Acts as a receptor for Ebolavirus, possibly through GAS6 binding to phosphatidyl-serine at the surface of virion envelope.

PTMs:

Monoubiquitinated upon GAS6-binding. A very small proportion of the receptor could be subjected to polyubiquitination in a very transient fashion.

Phosphorylated at tyrosine residues by autocatalysis, which activates kinase activity.

細胞の位置付け:

Cell membrane>Single-pass type I membrane protein.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
組織特異性:

Highly expressed in metastatic colon tumors. Expressed in primary colon tumors. Weakly expressed in normal colon tissue.

サブユニット構造:

Heterodimer and heterotetramer with ligand GAS6. 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.

タンパク質ファミリー:

Belongs to the protein kinase superfamily. Tyr protein kinase family. AXL/UFO subfamily.

研究領域

· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.

参考文献

1). Skipping of exon 10 in Axl pre-mRNA regulated by PTBP1 mediates invasion and metastasis process of liver cancer cells. Theranostics, 2020 (PubMed: 32483414) [IF=12.4]

Application: WB    Species: Human    Sample: HepG2 cells

Figure 2. Axl-S isoform has a more robust binding ability to Gas6 ligands. (A) Western-blot was used to detect the over-expression of Axl-L and Axl-S isoforms in HepG2 cells. (B) Left: Co-IP assay was used to detect the binding ability of Axl-L and Axl-S isoforms to Gas6 ligands; right: The relative amount of Gas6 protein bound in HepG2 cells over-expressing Axl-L or Axl-S. Data were analyzed using Student’s T-test. The control levels were set to a value of 1. Data are presented as mean ± S.D. (N=3). The “***” indicates “P<0.001” versus the pMXs-Axl-L group. (C) The effect of Axl-L and Axl-S over-expression on Axl and downstream AKT and ERK signaling pathway proteins was detected by Western-blot. Statistical analysis of the effects of over-expression of Axl isoforms on AKT and ERK signaling pathway proteins in HepG2 cells (D) and HCCLM3 cells (E). The relative expression of p-Axl represents the relative expression of p-Axl to Axl. The relative expression of p-AKT represents the relative expression of p-AKT to total Axl. The relative expression of p-ERK represents the relative expression of p-ERK to total ERK. The relative expression of total Axl, AKT, ERK represents the relative expression of total Axl, AKT, ERK to β-actin. Two-way ANOVA and Tukey’s post-hoc test was used to analyze the data. Data are presented as mean ± S.D. (N=3). The “*, **, ***” indicate “P<0.05, 0.01, 0.001” versus the control group, respectively. "#" represents P<0.05, "##" represents P<0.01, and "###" represents P<0.001. “#, ##, ###” indicates statistical difference between over-expressed Axl-L and Axl-S experimental groups.
2). Inhibitory effect of the novel tyrosine kinase inhibitor DCC-2036 on triple-negative breast cancer stem cells through AXL-KLF5 positive feedback loop. Cell death & disease, 2022 (PubMed: 36042208) [IF=9.0]

Application: WB    Species: Human    Sample: MDA-MB-231 cells

Fig. 3 DCC-2036 decreases the KLF5 expression through the inhibition of AXL. A DCC-2036 inhibited the activation and expression of AXL in a dose-dependent manner. MDA-MB-231 cells, HS-578T, and 4T1 cells were treated with indicated concentrations of DCC-2036 for 48 h, and then the phosphorated and total levels of AXL were detected by WB. B Gas6 increased the protein levels of p-AXL and KLF5. MDA-MB-231 cells, HS-578T, and 4T1 cells were treated with Gas6 (200 ng/ml) for 0, 30, 60, 90, 120 min. C The knockdown of AXL reduced the protein levels of p-AXL, AXL, and KLF5. MDA-MB-231 cells were transfected with human AXL siRNA or control siRNA (NC) for 48 h, and then the protein levels were tested by WB. D The mRNA levels of AXL, KLF5, and Nanog were quantified by q RT-PCR. The statistical significance was determined by Student’s t-test, **P 
3). Aberrant activation of AXL may drive progression of squamous cell carcinoma in CLL patients: a mechanistic study with clinical implications. British journal of cancer, 2024 (PubMed: 38886556) [IF=8.8]
4). Gas6 Attenuates Sepsis-Induced Tight Junction Injury and Vascular Endothelial Hyperpermeability via the Axl/NF-κB Signaling Pathway. Frontiers in Pharmacology, 2019 (PubMed: 31263416) [IF=5.6]

Application: WB    Species: mouse    Sample: MAECs

FIGURE 5 | TAM receptor expression in MAECs and Axl siRNA transfection efficiency. (A, C) Western blotting analysis of the expression of TAM receptors (Tyro3, Axl, and Mertk) in MACEs (n = 3, means ± SD). (B, D) Axl was activated when MAECs were incubated with Gas6; Western blotting was used to examine the Axl phosphorylation expression (n = 3, means ± SD, **P < 0.01 versus the control group).

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