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AB322001

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

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Rabbit Recombinant Multiclonal ALK antibody. Carrier free. Suitable for Dot, WB, IP and reacts with Synthetic peptide - Human, Human samples.

View Alternative Names

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

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Immunoprecipitation - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] - BSA and Azide free (AB322001)
  • IP

Supplier Data

Immunoprecipitation - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] - BSA and Azide free (AB322001)

This data was developed using ab322000, the same antibody clone in a different buffer formulation.

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] (<a href='/en-us/products/primary-antibodies/alk-phospho-y1278-y1507-y1604-antibody-rm1164-ab322000'>ab322000</a>) 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

Western blot - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] - BSA and Azide free (AB322001)
  • WB

Supplier Data

Western blot - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] - BSA and Azide free (AB322001)

This data was developed using ab322000, the same antibody clone in a different buffer formulation.

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] (<a href='/en-us/products/primary-antibodies/alk-phospho-y1278-y1507-y1604-antibody-rm1164-ab322000'>ab322000</a>) 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

Western blot - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] - BSA and Azide free (AB322001)
  • WB

Supplier Data

Western blot - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] - BSA and Azide free (AB322001)

This data was developed using ab322000, the same antibody clone in a different buffer formulation.

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] (<a href='/en-us/products/primary-antibodies/alk-phospho-y1278-y1507-y1604-antibody-rm1164-ab322000'>ab322000</a>) 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

Dot Blot - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] - BSA and Azide free (AB322001)
  • Dot

Supplier Data

Dot Blot - Anti-ALK (phospho Y1278 + Y1507 + Y1604) antibody [RM1164] - BSA and Azide free (AB322001)

This data was developed using ab322000, the same antibody clone in a different buffer formulation.

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] (<a href='/en-us/products/primary-antibodies/alk-phospho-y1278-y1507-y1604-antibody-rm1164-ab322000'>ab322000</a>) 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

  • Unconjugated

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

Key facts

Host species

Rabbit

Clonality

Multiclonal

Clone number

RM1164

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

IP, WB, Dot

applications

Immunogen

This product was produced with the following immunogens:

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

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

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Reactivity data

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Product details

ab322001 is the carrier-free version of ab322000.

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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
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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:
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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).
See full target information ALK

Additional targets

ALK tyrosine kinase receptor phospho Y1278,ALK phospho Y1507,ALK
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Product promise

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