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Rabbit Recombinant Monoclonal ALK phospho Y1278 antibody. Suitable for WB, IP, Dot and reacts with Human samples.

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Images

Immunoprecipitation - Anti-ALK(pY1278) antibody [EPR26945-22] (AB305099), expandable thumbnail
  • Dot Blot - Anti-ALK(pY1278) antibody [EPR26945-22] (AB305099), expandable thumbnail
  • Western blot - Anti-ALK(pY1278) antibody [EPR26945-22] (AB305099), expandable thumbnail
  • Western blot - Anti-ALK(pY1278) antibody [EPR26945-22] (AB305099), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBIHC-PICC/IFFlow Cyt (Intra)IPDot
Human
Tested
Not recommended
Not recommended
Not recommended
Tested
Tested
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species
Mouse
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species
Rat
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Human, Mouse, Rat
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/30
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Target data

Function

Neuronal receptor tyrosine kinase that is essentially and transiently expressed in specific regions of the central and peripheral nervous systems and plays an important role in the genesis and differentiation of the nervous system (PubMed:11121404, PubMed:11387242, PubMed:16317043, PubMed:17274988, PubMed:30061385, PubMed:34646012, PubMed:34819673). Also acts as a key thinness protein involved in the resistance to weight gain: in hypothalamic neurons, controls energy expenditure acting as a negative regulator of white adipose tissue lipolysis and sympathetic tone to fine-tune energy homeostasis (By similarity). Following activation by ALKAL2 ligand at the cell surface, transduces an extracellular signal into an intracellular response (PubMed:30061385, PubMed:33411331, PubMed:34646012, PubMed:34819673). In contrast, ALKAL1 is not a potent physiological ligand for ALK (PubMed:34646012). Ligand-binding to the extracellular domain induces tyrosine kinase activation, leading to activation of the mitogen-activated protein kinase (MAPK) pathway (PubMed:34819673). Phosphorylates almost exclusively at the first tyrosine of the Y-x-x-x-Y-Y motif (PubMed:15226403, PubMed:16878150). Induces tyrosine phosphorylation of CBL, FRS2, IRS1 and SHC1, as well as of the MAP kinases MAPK1/ERK2 and MAPK3/ERK1 (PubMed:15226403, PubMed:16878150). ALK activation may also be regulated by pleiotrophin (PTN) and midkine (MDK) (PubMed:11278720, PubMed:11809760, PubMed:12107166, PubMed:12122009). PTN-binding induces MAPK pathway activation, which is important for the anti-apoptotic signaling of PTN and regulation of cell proliferation (PubMed:11278720, PubMed:11809760, PubMed:12107166). MDK-binding induces phosphorylation of the ALK target insulin receptor substrate (IRS1), activates mitogen-activated protein kinases (MAPKs) and PI3-kinase, resulting also in cell proliferation induction (PubMed:12122009). Drives NF-kappa-B activation, probably through IRS1 and the activation of the AKT serine/threonine kinase (PubMed:15226403, PubMed:16878150). Recruitment of IRS1 to activated ALK and the activation of NF-kappa-B are essential for the autocrine growth and survival signaling of MDK (PubMed:15226403, PubMed:16878150).

Alternative names

CD246, ALK tyrosine kinase receptor, Anaplastic lymphoma kinase, ALK

Recommended products

Rabbit Recombinant Monoclonal ALK phospho Y1278 antibody. Suitable for WB, IP, Dot and reacts with Human samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR26945-22
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

The Anaplastic Lymphoma Kinase commonly referred to as ALK is a receptor tyrosine kinase known for its role in cell signaling. It is sometimes called CD246. ALK has a molecular mass of approximately 180 kDa. This protein finds expression predominantly in the central and peripheral nervous system with prominence in neural tissue during development. The ALK protein belongs to the insulin receptor superfamily exhibiting kinase activity that promotes signal transduction processes associated with growth and differentiation.

Biological function summary

ALK influences cell growth survival and differentiation playing a significant role during the development of the nervous system. As part of its biological activity the ALK protein can become a component of larger signaling complexes participating as an important activator or mediator. Evidence suggests ALK's involvement in neuronal differentiation and synaptogenesis. Its activity impacts intracellular pathways consequently modulating biological processes relevant to neural tissue and oncogenesis.

Pathways

ALK acts within the MAPK and PI3K-AKT signaling pathways. These pathways enable the transduction of signals from the cell surface to the nucleus influencing cellular outcomes such as proliferation and survival. In these pathways ALK interacts with various other proteins including GRB2 and PI3K which further facilitate downstream signaling. Proper functioning of these pathways is essential to maintaining cellular homeostasis and development.

Associated diseases and disorders

ALK has a significant relationship with certain types of cancer such as non-small cell lung carcinoma and anaplastic large cell lymphoma. Genetic alterations in ALK such as translocations or mutations can lead to the uncontrolled activation of its kinase activity resulting in oncogenic transformation. In these contexts the ALK protein often interacts with EML4 in lung cancer through a fusion forming an oncogenic driver. Targeting ALK with anti-ALK antibodies or small molecule inhibitors has emerged as a therapeutic strategy to manage these malignancies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

4 product images

  • Immunoprecipitation - Anti-ALK(pY1278) antibody [EPR26945-22] (ab305099), expandable thumbnail

    Immunoprecipitation - Anti-ALK(pY1278) antibody [EPR26945-22] (ab305099)

    ALK(pY1278) was immunoprecipitated from 0.35 mg KARPAS-299 (human T cell lymphoma cell) treated with 50uM pervanadate for 30min, whole cell lysate 10ug with ab305099 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305099 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: KARPAS-299 (human T cell lymphoma cell) treated with 50uM pervanadate for 30min, whole cell lysate 10ug
    Lane 2: abAB305099 IP in KARPAS-299 treated with 50uM pervanadate for 30min, whole cell lysate
    Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab305099 in KARPAS-299 treated with 50uM pervanadate for 30min, whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 84 seconds

    All lanes: Immunoprecipitation - Anti-ALK(pY1278) antibody [EPR26945-22] (ab305099) at 1/1000 dilution

    Lane 1: KARPAS-299 (human T cell lymphoma cell) treated with 50uM pervanadate for 30min, whole cell lysate 10ug

    Lane 2: abAB305099 IP in KARPAS-299 treated with 50uM pervanadate for 30min, whole cell lysate

    Secondary

    All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

    Predicted band size: 80 kDa

    Observed band size: 80 kDa

    Exposure time: 84s

  • Dot Blot - Anti-ALK(pY1278) antibody [EPR26945-22] (ab305099), expandable thumbnail

    Dot Blot - Anti-ALK(pY1278) antibody [EPR26945-22] (ab305099)

    Dot blot analysis of ALK(pY1278) using ab305099 at 1:1000 (0.476 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.

    Exposure time: 3 minutes
    Blocking and diluting buffer and concentration: 5% NFDM/TBST

  • Western blot - Anti-ALK(pY1278) antibody [EPR26945-22] (ab305099), expandable thumbnail

    Western blot - Anti-ALK(pY1278) antibody [EPR26945-22] (ab305099)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST
    KARPAS-299 cells carry the NPM-ALK fusion (PMID: 32193476, 9736036).

    Phosphorylated Tyr338 of NPM-ALK is equivalent to Tyr1278 of full length ALK.

    Bands around 300 kDa may be due to protein aggregation (PMID: 9121481).

    Lysates were prepared from fresh material and used for Western blotting immediately to minimize protein degradation.
    Exposure time: 81 seconds

    All lanes: Western blot - Anti-ALK(pY1278) antibody [EPR26945-22] (ab305099) at 1/1000 dilution

    Lane 1: Untreated KARPAS-299 (human anaplastic large cell lymphoma) whole cell lysate at 20 µg

    Lane 2: KARPAS-299 treated with 1 uM Crizotinib for 1 hour whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 176 kDa

    Observed band size: 300 kDa, 80 kDa

  • Western blot - Anti-ALK(pY1278) antibody [EPR26945-22] (ab305099), expandable thumbnail

    Western blot - Anti-ALK(pY1278) antibody [EPR26945-22] (ab305099)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST
    KARPAS-299 cells carry the NPM-ALK fusion (PMID: 32193476, 9736036).

    Phosphorylated Tyr338 of NPM-ALK is equivalent to Tyr1278 of full length ALK.

    Bands around 300 kDa may be due to protein aggregation (PMID: 9121481).

    Lysates were prepared from fresh material and used for Western blotting immediately to minimize protein degradation.
    Exposure time: 81 seconds

    All lanes: Western blot - Anti-ALK(pY1278) antibody [EPR26945-22] (ab305099) at 1/1000 dilution

    Lane 1: KARPAS-299 (human anaplastic large cell lymphoma), whole cell lysate (Untreated membrane) at 20 µg

    Lane 2: KARPAS-299 treated with 50 uM pervanadate for 30 minutes, whole cell lysate (Untreated membrane) at 20 µg

    Lane 3: KARPAS-299 whole cell lysate (phosphatase treated membrane) at 20 µg

    Lane 4: KARPAS-299 treated with 50 uM pervanadate for 30 minutes, whole cell lysate (phosphatase treated membrane) at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 176 kDa

    Observed band size: 80 kDa, 300 kDa

    Exposure time: 81s

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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