AB322000

Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164]

20ul selling size
Recombinant
RabMAb
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Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] is a Rabbit Recombinant Multiclonal antibody that is used in ALK (phospho Y1278 + Y1507 + Y1604) Dot Blot, IP, Western Blot. Suitable for Human samples.

ALK (CD246) is a receptor tyrosine kinase which plays a role in cell communication and development of the nervous system. Phosphorylated ALK activates multiple downstream signal transduction pathways, including JAK-STAT and MAPK-ERK. ALK has been implicated in the pathogenecis of several types of cancer, including non-Hodgkin lumphoma, lung adenocarcinoma and anaplastic large-cell lymphoma.

View Alternative Names

CD246, ALK tyrosine kinase receptor, Anaplastic lymphoma kinase

4 Images

Supplier Data

Immunoprecipitation - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] (AB322000)

ALK (phospho Y1278 + Y1507 + Y1604) was immunoprecipitated from 0.35 mg KARPAS-299 (human T cell lymphoma cell) whole cell lysate with ab322000 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322000 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : KARPAS-299 (human T cell lymphoma cell) whole cell lysate
Lane 2 : ab322000 IP in KARPAS-299 (human T cell lymphoma cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab322000 in KARPAS-299 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] (ab322000) at 1/30 dilution

All lanes:

KARPAS-299 (human T cell lymphoma cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 3s

Supplier Data

Western blot - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] (AB322000)

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

KARPAS-299 cells carry the NPM-ALK fusion (PMID : 32193476, PMID : 9736036).

Phosphorylated Tyr338/ Tyr567/ Tyr664 of NPM-ALK is equivalent to Tyr1278/ Tyr1507/ Tyr1604 of full length ALK (PMID : 19887368).

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-ALK antibody [SP144] (ab183332) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] (ab322000) 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, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 80 kDa,300 kDa,36 kDa

false

Exposure time: 180s

Supplier Data

Western blot - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] (AB322000)

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

KARPAS-299 cells carry the NPM-ALK fusion (PMID : 32193476, PMID : 9736036).

Phosphorylated Tyr338/ Tyr567/ Tyr664 of NPM-ALK is equivalent to Tyr1278/ Tyr1507/ Tyr1604 of full length ALK (PMID : 19887368).

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.

In Western blot, Anti-ALK antibody [SP144] (ab183332) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] (ab322000) 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 whole cell lysate (phosphatase treated membrane) at 20 µg

Lane 3:

KARPAS-299 treated with 50 uM pervanadate for 30 minutes, whole cell lysate (Untreated 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 80 kDa,300 kDa

false

Exposure time: 59s

Supplier Data

Dot Blot - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] (AB322000)

Dot blot analysis of ALK (phospho Y1278 + Y1507 + Y1604) using ab322000 at 1 : 1000 dilution (0.476 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : Human ALK tyrosine kinase receptor (phospho Y1604) peptide (aa1600-1609)
Lane2 : Human ALK tyrosine kinase receptor peptide (aa1599-1610)
Lane3 : Human ALK tyrosine kinase receptor (phospho Y1507) peptide (aa1501-1516)
Lane4 : Human ALK tyrosine kinase receptor peptide (aa1500-1516)
Lane5 : Human ALK tyrosine kinase receptor (phospho Y1278) peptide (aa1272-1284)
Lane6 : Human ALK tyrosine kinase receptor peptide (aa1272-1284)
Lane7 : Human ALK tyrosine kinase receptor (phospho Y1282) peptide (aa1272-1286)
Lane8 : Human ALK tyrosine kinase receptor (phospho Y1283) peptide (aa1272-1286)
Lane9 : Human ALK tyrosine kinase receptor (phospho Y1282+Y1283) peptide (aa1272-1286)

Exposure time : 180 seconds.

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

All lanes:

Dot Blot - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] (ab322000) at 1/1000 dilution

Lane 1:

Human ALK tyrosine kinase receptor (phospho Y1604) peptide (aa1600-1609)

Lane 2:

Human ALK tyrosine kinase receptor peptide (aa1599-1610)

Lane 3:

Human ALK tyrosine kinase receptor (phospho Y1507) peptide (aa1501-1516)

Lane 4:

Human ALK tyrosine kinase receptor peptide (aa1500-1516)

Lane 5:

Human ALK tyrosine kinase receptor (phospho Y1278) peptide (aa1272-1284)

Lane 6:

Human ALK tyrosine kinase receptor peptide (aa1272-1284)

Lane 7:

Human ALK tyrosine kinase receptor (phospho Y1282) peptide (aa1272-1286)

Lane 8:

Human ALK tyrosine kinase receptor (phospho Y1283) peptide (aa1272-1286)

Lane 9:

Human ALK tyrosine kinase receptor (phospho Y1282+Y1283) peptide (aa1272-1286)

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 180s

Carrier free

Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] - BSA and Azide free

Key facts

Host species

Rabbit

Clonality

Multiclonal

Clone number

RM1164

Isotype

IgG

Carrier free

Reacts with

Human

Applications

WB, IP, Dot

applications

Immunogen

This product was produced with the following immunogens:

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/30", "IP-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" }, "Mouse": { "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" }, "Rat": { "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" }, "Synthetic peptide - Human": { "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "1/1000", "Dot-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" } } }

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Product details

What are recombinant multiclonals?
Recombinant multiclonals are a mixture of recombinant antibodies co-expressed from a library of heavy and light chains. They offer several advantages including:

- The sensitivity of polyclonal antibodies by recognising multiple epitopes
- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

View our range of recombinant multiclonal antibodies.

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

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

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

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

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Supplementary information

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

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.

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.

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Product protocols

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

Visit the General protocols
Visit the Troubleshooting

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Target data

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). May function as regulator of gastric epithelial differentiation (By similarity).

See full target information ALK tyrosine kinase receptor phospho Y1278 + Y1507 + Y1604

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164]

Immunoprecipitation - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] (AB322000)

Western blot - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] (AB322000)

Western blot - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] (AB322000)

Dot Blot - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] (AB322000)

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Complementary Products

Primary Antibodies

Secondary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage duration

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Aliquoting information

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Product promise