Human Phospho-Axl (Y779) Antibody

(5 citations)
(3 Reviews)
  
  • Species Reactivity
    Human
  • Specificity
    Detects human and mouse Axl when phosphorylated at Y779.
  • Source
    Polyclonal Rabbit IgG
  • Purification
    Antigen and protein A Affinity-purified
  • Immunogen
    Phosphopeptide containing human Axl Y779 site
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Immunohistochemistry
Phospho-Axl (Y779) in Human Stomach Cancer Tissue. Axl phosphorylated at site Y779 was detected in immersion fixed paraffin-embedded sections of human stomach cancer tissue using Human Phospho-Axl (Y779) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2228) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). Specific labeling was localized to the cytoplasm of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human Phospho-Axl (Y779) by Western Blot. Western blot shows lysates of A172 human glioblastoma cell line untreated (-) or treated (+) with WI38 human lung cell line conditioned media (WI38 CM) for 15 minutes. PVDF membrane was probed with 1 µg/mL of Human Phospho‑Axl (Y779) Antigen Affinity‑purified Polyclonal Antibody, followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho‑Axl (Y779) at approximately 140 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
Phospho-Axl (Y779) in A172 Human Cell Line. Axl phosphorylated at Y779 (panels B, D) and total Axl (panels A, C) were assessed in immersion fixed A172 human glioblastoma cells incubated with (panels C, D) or without (panels A, B) pervanadate. Phospho-Axl was detected using Rabbit Anti-Human Phospho-Axl (Y779) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2228) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red, panels B, D); Catalog # NL004) and counterstained using DAPI (blue). Total Axl was detected using Goat Anti-Human Axl Antigen Affinity-purified Polyclonal Antibody (Catalog # AF154). Cells were stained using the NorthernLights™ 493-conjugated Anti-Goat IgG Secondary Antibody (green, panels A, C); Catalog # NL003). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Axl

Axl (Ufo, Ark), Dtk (Sky, Tyro3, Rse, Brt), and Mer (human and mouse homologues of chicken c-Eyk) constitute a subfamily of the receptor tyrosine kinases (1, 2). The extracellular domains of these proteins contain two Ig-like motifs and two fibronectin type III motifs. This characteristic topology is also found in neural cell adhesion molecules and in receptor tyrosine phosphatases. The human Axl cDNA encodes an 887 amino acid (aa) precursor that includes an 18 aa signal sequence, a 426 aa extracellular domain, a 21 aa transmembrane segment, and a 422 aa cytoplasmic domain. The extracellular domains of human and mouse Axl share 81% aa sequence identity. A short alternately spliced form of human Axl is distinguished by a 9 aa deletion in the extracellular juxtamembrane region. These receptors bind the vitamin K‑dependent protein growth arrest specific gene 6 (Gas6) which is structurally related to the anticoagulation factor protein S. Binding of Gas6 induces receptor autophosphorylation and downstream signaling pathways that can lead to cell proliferation, migration, or the prevention of apoptosis (3). This family of tyrosine kinase receptors is involved in hematopoiesis, embryonic development, tumorigenesis, and regulation of testicular functions.

  • References:
    1. Yanagita, M. (2004) Curr. Opin. Nephrol. Hypertens. 13:465.
    2. Nagata, K. et al. (1996) J. Biol. Chem. 22:30022.
    3. Holland, S. et al. (2005) Canc. Res. 65:9294.
  • Long Name:
    Axl Receptor Tyrosine Kinase
  • Entrez Gene IDs:
    558 (Human); 26362 (Mouse)
  • Alternate Names:
    Ark; AXL oncogene; AXL receptor tyrosine kinase; AXL transforming sequence/gene; Axl; EC 2.7.10; EC 2.7.10.1; JTK11; tyrosine-protein kinase receptor UFO; Ufo; UFOoncogene AXL
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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Species
Applications
Sample Type
  1. Tropomyosin isoform Tpm2.1 regulates collective and amoeboid cell migration and cell aggregation in breast epithelial cells
    Authors: H Shin, D Kim, DM Helfman
    Oncotarget, 2017;0(0):.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. Receptor tyrosine kinase Axl is required for resistance of leukemic cells to FLT3-targeted therapy in acute myeloid leukemia.
    Authors: Park I, Mundy-Bosse B, Whitman S, Zhang X, Warner S, Bearss D, Blum W, Marcucci G, Caligiuri M
    Leukemia, 2015;29(12):2382-9.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. Identification of Axl as a downstream effector of TGF-beta1 during Langerhans cell differentiation and epidermal homeostasis.
    J. Exp. Med., 2012;209(11):2033-47.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Frozen
  4. The novel receptor tyrosine kinase Axl is constitutively active in B-cell chronic lymphocytic leukemia and acts as a docking site of nonreceptor kinases: implications for therapy.
    Authors: Ghosh AK, Secreto C, Boysen J, Sassoon T, Shanafelt TD, Mukhopadhyay D, Kay NE
    Blood, 2011;117(6):1928-37.
    Species: Human
    Sample Type: Cell Lysates
    Application: IP
  5. Growth arrest-specific protein 6 deficiency impairs liver tissue repair after acute toxic hepatitis in mice.
    Authors: Lafdil F, Chobert MN, Deveaux V, Zafrani ES, Mavier P, Nakano T, Laperche Y, Brouillet A
    J. Hepatol., 2009;51(1):55-66.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
Cell and Tissue Staining Kits
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CTS005 3
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Immunohistochemistry Reagents
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Normal Rabbit IgG Control

Ctrl AB-105-C 43  
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Ctrl, CyTOF-ready MAB1050  
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Secondary Antibodies
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Rabbit IgG HRP-conjugated Antibody

WB HAF008 23  
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Donkey Anti-Rabbit IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL004 10
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Donkey Anti-Rabbit IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL005 2
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Rabbit IgG APC-conjugated Antibody

Flow F0111 1  
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Rabbit IgG PE-conjugated Antibody

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Rabbit IgG Fluorescein-conjugated Antibody

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Donkey Anti-Rabbit IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL006 4
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Goat Anti-Rabbit IgG Biotinylated Antibody

WB BAF008 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Rabbit IgG Antibody

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Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC003  
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Donkey Anti-Rabbit IgG (H+L) Antibody

D-301-C-ABS2
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Average Rating: 3.7 (Based on 3 reviews)

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We have 2 reviews tested in 2 species: Human, Mouse.
We have 3 review tested in 1 application: Western Blot.

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Images Ratings Applications Species Reviewed By Date Details
  Very Good
 WB Mouse Anonymous 06/23/2017
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Summary

ApplicationWestern Blot
Sample TestedBrain tissue
SpeciesMouse
Western Blot Human Phospho-Axl (Y779) Antibody AF2228
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Very Good
 WB Anonymous 10/06/2016
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Western Blot Human Phospho-Axl (Y779) Antibody AF2228
Western Blot: Human Phospho-Axl (Y779) Antibody [AF2228]

Summary

ApplicationWestern Blot
Sample TestedMDA-MB-231 human breast cancer cell line,PC-3 human prostate cancer cell line,DU145 human prostate carcinoma cell line,LNCaP human prostate cancer cell line,S521,S156,U-87 MG human glioblastoma/astrocytoma cell line,U-118-MG human glioblastoma/astrocytoma cell line,RKO,KHT,SCC-7
  Average
 WB Human Anonymous 04/25/2016
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Summary

ApplicationWestern Blot
Sample TestedMCF 10A human breast epithelial cell line,mcf10aDCIS.com,MDA-MB-231 human breast cancer cell line
SpeciesHuman

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