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Human Phospho-Axl (Y779) Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB6965

R&D Systems, part of Bio-Techne

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

CyTOF-ready, Immunocytochemistry, Intracellular Staining by Flow Cytometry, Simple Western, Western Blot

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 713610

Product Specifications

Immunogen

Phosphopeptide containing the human Axl Y779 site

Specificity

Detects human Phospho-Axl (Y779) in direct ELISAs and Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human Phospho-Axl (Y779) Antibody

Detection of Human Phospho-Axl (Y779) antibody by Western Blot.

Detection of Human Phospho-Axl (Y779) by Western Blot.

Western blot shows lysates of A172 human glioblastoma cell line untreated (-) or treated (+) with 1 mM Pervanadate (PV) for 30 minutes. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-Human Phospho-Axl (Y779) Monoclonal Antibody (Catalog # MAB6965) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). 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.
Phospho-Axl (Y779) antibody in A172 Human Cell Line by Immunocytochemistry (ICC).

Phospho-Axl (Y779) in A172 Human Cell Line.

Axl phosphorylated at Y779 and total Axl were assessed in immersion fixed A172 human glioblastoma cells incubated with (upper panel) or without (lower panel) the phosphatase inhibitor pervanadate at 100 µM for 5 minutes. Phospho-Axl was detected using Mouse Anti-Human Phospho-Axl (Y779) Monoclonal Antibody (Catalog # MAB6965) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) 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; Catalog # NL003). Specific staining was localized to cytoplasm and cell surfaces. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Phospho-Axl (Y779) antibody in A172 Human Cell Line antibody by Flow Cytometry.

Detection of Phospho-Axl (Y779) in A172 Human Cell Line by Flow Cytometry.

A172 human glioblastoma cell line was unstimulated (open histogram) or treated with 100 µM pervanadate for 5 minutes, then stained with Mouse Anti-Human Phospho-Axl (Y779) Monoclonal Antibody (Catalog # MAB6965, filled histo-gram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with Triton® X-100.

Applications for Human Phospho-Axl (Y779) Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed A172 human glioblastoma cell line treated with or without pervanadate

Intracellular Staining by Flow Cytometry

2.5 µg/106 cells
Sample: Pervanadate-treated A172 human glioblastoma cell line fixed with paraformaldehyde and permeabilized with Triton® X-100

Simple Western

4 µg/mL
Sample: A172 human glioblastoma cell line treated with Pervanadate (PV)

Western Blot

0.2 µg/mL
Sample: A172 human glioblastoma cell line treated with Pervanadate (PV)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
Size / Price
Qty
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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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. Phosphorylation of Tyrosine 779 provides a docking site for p85 subunits of PI 3-Kinase (4).

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.
  4. Weinger, J.G. et al. (2008) J. Neurochem. 106:134.

Long Name

Axl Receptor Tyrosine Kinase

Alternate Names

AI323647, ARK, JTK11, Tyro7, UFO

Entrez Gene IDs

558 (Human); 26362 (Mouse); 102126361 (Cynomolgus Monkey)

Gene Symbol

AXL

Additional Axl Products

Product Documents for Human Phospho-Axl (Y779) Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Phospho-Axl (Y779) Antibody

For research use only

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