Anti-ALK antibody [EPR29054-38] (ab323643)

Rabbit Recombinant Monoclonal ALK antibody. Suitable for IHC-P, WB and reacts with Human, Mouse samples.

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Images

Western blot - Anti-ALK antibody [EPR29054-38] (AB323643), 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

	IHC-P	WB
Human	Tested	Tested
Mouse	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/200	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/100 - 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -

Target data

See full target information ALK

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

Rabbit Recombinant Monoclonal ALK antibody. Suitable for IHC-P, WB and reacts with Human, Mouse samples.

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.
Clone number: EPR29054-38
Purification technique: Affinity purification Protein A
Specificity: ALK expression is low in healthy tissue and can vary significantly in solid tumors. ALK expression was not observed in healthy or cancer tissue microarrays tested in IHC-P
Concentration

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

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

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.

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.

9 product images

Western blot - Anti-ALK antibody [EPR29054-38] (ab323643)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: U-937 (PMID: 9736036).
KARPAS-299 cells carry the NPM-ALK fusion (PMID: 9736036, PMID: 11310834). Bands higher than 250 KDa are likely representing oligomers of NPM-ALK (PMID: 23610619).
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 9736036, PMID: 11310834, 9121481).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-ALK antibody [EPR29054-38] (ab323643) at 1/1000 dilution
Lane 1: KARPAS-299 (human T cell lymphoma cell) whole cell lysate at 20 µg
Lane 2: U-937 (human histiocytic lymphoma monocyte) 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
Observed band size: 80 kDa, 36 kDa
Exposure time: 10s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALK antibody [EPR29054-38] (ab323643)
Immunohistochemical analysis of paraffin-embedded Adult mouse kidney tissue labeling ALK with ab323643 at 1/100 (5.26 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Negative control: No staining on Adult mouse kidney.

The section was incubated with ab323643 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Western blot - Anti-ALK antibody [EPR29054-38] (ab323643)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: RAW 264.7 (30541790), kidney (PMID: 9053841).
Low expression: hippocampus (PMID: 37291945, PMID: 9053841).
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 9736036, PMID: 11310834, 9121481).
The identity of the lower MW band at approximately 37 kDa (in lanes 4-5) is unknown.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-ALK antibody [EPR29054-38] (ab323643) at 1/1000 dilution
Lane 1: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg
Lane 2: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Lane 3: Mouse P5 brain tissue lysate at 20 µg
Lane 4: Mouse kidney tissue lysate at 20 µg
Lane 5: Mouse hippocampus tissue 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
Observed band size: 250 kDa, 36 kDa
Exposure time: 180s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALK antibody [EPR29054-38] (ab323643)
Immunohistochemical analysis of paraffin-embedded Adult mouse cerebral cortex tissue labeling ALK with ab323643 at 1/100 (5.26 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Low expression tissue: Nearly no staining on adult mouse cerebral cortex.

The section was incubated with ab323643 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Western blot - Anti-ALK antibody [EPR29054-38] (ab323643)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: Raji (PMID: 30541790), Jurkat (PMID: 10793082).
Full-length ALK is approximately 250 kDa. Band around 130kDa might be cleavage fragment(PMID: 30541790).
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 9736036, PMID: 11310834, 9121481).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-ALK antibody [EPR29054-38] (ab323643) at 1/1000 dilution
Lane 1: IMR-32 (human neuroblastoma neuroblast) whole cell lysate at 20 µg
Lane 2: Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg
Lane 3: Jurkat (human T cell leukemia T lymphocyte from peripheral blood) 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
Observed band size: 250 kDa, 36 kDa
Exposure time: 180s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALK antibody [EPR29054-38] (ab323643)
Immunohistochemical analysis of paraffin-embedded E15.5 Mouse embryo tissue labeling ALK with ab323643 at 1/100 (5.26 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on brain and peripheral nervous system of E15.5 mouse embryo.

The section was incubated with ab323643 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALK antibody [EPR29054-38] (ab323643)
Immunohistochemical analysis of paraffin-embedded (A) Neuro-2a (mouse neuroblastoma neuroblast) cell pellet. (B) RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cell pellet. tissue labeling ALK with ab323643 at 1/100 (5.26 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Positive staining on (A) Neuro-2a cell pellet, no staining on (B) RAW 264.7 cell pellet.

The section was incubated with ab323643 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALK antibody [EPR29054-38] (ab323643)
Immunohistochemical analysis of paraffin-embedded (A) Mouse PDX model of human anaplastic large cell lymphoma. (B) Control mouse PDX model. tissue labeling ALK with ab323643 at 1/2000 (0.263 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on (A) Mouse PDX model of human anaplastic large cell lymphoma. No staining on (B) Control mouse PDX model.

The section was incubated with ab323643 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALK antibody [EPR29054-38] (ab323643)
Immunohistochemical analysis of paraffin-embedded (A) KARPAS-299 (human T cell lymphoma cell) cell pellet. (B) U-937 (human histiocytic lymphoma monocyte) cell pellet. tissue labeling ALK with ab323643 at 1/200 (2.63 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Positive staining on (A) KARPAS-299 cell pellet, no staining on (B) U-937 cell pellet.

The section was incubated with ab323643 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins