Immunohistochemistry analysis of ALK 400 showing staining in the cytoplasm of paraffin-embedded human skin tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a ALK 400 Mouse Monoclonal Antibody (354300) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
|Tested species reactivity||Human|
|Published species reactivity||Human, Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant protein derived from the human ALK protein.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:50|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
The 2;5 chromosomal translocation is frequently associated with anaplastic large cell lymphomas. The translocation creates a fusion gene consisting of the ALK gene and the nucleophosmin gene: the 3' half of ALK, derived from chromosome 2, is fused to the 5' portion of NPM from chromosome 5. A recent study shows that the product of the NPM-ALK fusion gene is oncogenic. The deduced amino acid sequences reveal that ALK is a novel receptor protein-tyrosine kinase having a putative transmembrane domain and an extracellular domain. These sequences are absent in the product of the transforming NPM-ALK gene. ALK shows the greatest sequence similarity to LTK. ALK plays an important role in the development of the brain and exerts its effects on specific neurons in the nervous system.
IP-MS enrichment of ALK (LFQ intensity): ALK was enriched 416-fold from SR lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Part No. 90409) and ALK antibody (Part No. 35-4300). The STRING database (www.string-db.org) was used to identify the protein interactor list. See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Excess of NPM-ALK oncogenic signaling promotes cellular apoptosis and drug dependency.
35-4300 was used in immunocytochemistry, immunohistochemistry, and western blot to study NPM-ALK-amplified cell lines resistant to ALK tyrosine kinase inhibitors
|Ceccon M,Merlo ME,Mologni L,Poggio T,Varesio LM,Menotti M,Bombelli S,Rigolio R,Manazza AD,Di Giacomo F,Ambrogio C,Giudici G,Casati C,Mastini C,Compagno M,Turner SD,Gambacorti-Passerini C,Chiarle R,Voena C||Oncogene (35:3854)||2016|
|Not Applicable||Not Cited||
Entrectinib, a Pan-TRK, ROS1, and ALK Inhibitor with Activity in Multiple Molecularly Defined Cancer Indications.
35-4300 was used in western blot to characterize multiple molecularly defined cancer indications by studying ROS1, ALK inhibitor, Entrectinib, a Pan-TRK activity
|Ardini E,Menichincheri M,Banfi P,Bosotti R,De Ponti C,Pulci R,Ballinari D,Ciomei M,Texido G,Degrassi A,Avanzi N,Amboldi N,Saccardo MB,Casero D,Orsini P,Bandiera T,Mologni L,Anderson D,Wei G,Harris J,Vernier JM,Li G,Felder E,Donati D,Isacchi A,Pesenti E,Magnaghi P,Galvani A||Molecular cancer therapeutics (15:628)||2016|
CEP-28122, a highly potent and selective orally active inhibitor of anaplastic lymphoma kinase with antitumor activity in experimental models of human cancers.
35-4300 was used in western blot to identify and characterize an anaplastic lymphoma kinase inhibitor.
|Cheng M,Quail MR,Gingrich DE,Ott GR,Lu L,Wan W,Albom MS,Angeles TS,Aimone LD,Cristofani F,Machiorlatti R,Abele C,Ator MA,Dorsey BD,Inghirami G,Ruggeri BA||Molecular cancer therapeutics (11:670)||2012|
|Human||Not Cited||NPM-ALK inhibits the p53 tumor suppressor pathway in an MDM2 and JNK-dependent manner.||Cui YX,Kerby A,McDuff FK,Ye H,Turner SD||Blood (113:5217)||2009|
||NPM-ALK inhibits the p53 tumor suppressor pathway in an MDM2 and JNK-dependent manner.||Cui YX,Kerby A,McDuff FK,Ye H,Turner SD||Blood (113:5217)||2009|