c-Kit protein (Active) (ab83580)
Constituents: 10% Trehalose, 1% Human serum albumin
Our Abpromise guarantee covers the use of ab83580 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
SDS-PAGE: Use at an assay dependent dilution.
Under reducing conditions ab83580 migrates as a broad band between 65 and 100 kDa on SDS-PAGE due to post-translational modifications, in particular glycosylation. This compares with unmodified c-Kit polypeptide that has a predicted monomeric molecular mass of 55.9 kDa. ab83580 consists of approximately 15-45% carbohydrate by weight.
ab83580 separates into a number of glycoforms with a pI between 4 and 7 on 2D PAGE due to post-translational modifications, in particular glycosylation. This compares with the unmodified c-Kit that has a predicted pI of 6.1.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
- C Kitc-KitCD117
- KITKIT oncogeneKIT_HUMANMast cell growth factor receptorMast/stem cell growth factor receptorMast/stem cell growth factor receptor Kitp145 c-kitPBTPiebald trait proteinProto oncogene c KitProto oncogene tyrosine protein kinase KitProto-oncogene c-KitSCF ReceptorSCFRsoluble KIT variant 1Steel Factor ReceptorStem cell factor receptortyrosine protein kinase KitTyrosine-protein kinase Kitv kit Hardy Zuckerman 4 feline sarcoma viral oncogene homologv kit Hardy Zuckerman 4 feline sarcoma viral oncogene like proteinv-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog
Defects in KIT are a cause of gastrointestinal stromal tumor (GIST) [MIM:606764].
Defects in KIT have been associated with testicular germ cell tumor (TGCT) [MIM:273300]. A common solid malignancy in males. Germ cell tumors of the testis constitute 95% of all testicular neoplasms.
Defects in KIT are a cause of acute myelogenous leukemia (AML) [MIM:601626]. AML is a malignant disease in which hematopoietic precursors are arrested in an early stage of development. Note=Somatic mutations that lead to constitutive activation of KIT are detected in AML patients. These mutations fall into two classes, the most common being in-frame internal tandem duplications of variable length in the juxtamembrane region that disrupt the normal regulation of the kinase activity. Likewise, point mutations in the kinase domain can result in a constitutively activated kinase.
Contains 5 Ig-like C2-type (immunoglobulin-like) domains.
Contains 1 protein kinase domain.
modificationsUbiquitinated by SOCS6. KIT is rapidly ubiquitinated after autophosphorylation induced by KITLG/SCF binding, leading to internalization and degradation.
Autophosphorylated on tyrosine residues. KITLG/SCF binding enhances autophosphorylation. Isoform 1 shows low levels of tyrosine phosphorylation in the absence of added KITLG/SCF (in vitro). Kinase activity is down-regulated by phosphorylation on serine residues by protein kinase C family members. Phosphorylation at Tyr-568 is required for interaction with PTPN11/SHP-2, CRK (isoform Crk-II) and members of the SRC tyrosine-protein kinase family. Phosphorylation at Tyr-570 is required for interaction with PTPN6/SHP-1. Phosphorylation at Tyr-703, Tyr-823 and Tyr-936 is important for interaction with GRB2. Phosphorylation at Tyr-721 is important for interaction with PIK3R1. Phosphorylation at Tyr-823 and Tyr-936 is important for interaction with GRB7.
c-Kit protein (Active) images
Densitometry of protein isoforms visualised by 2-DE. The densitometry scan demonstrates that the purified human cell expressed protein exists in multiple glycoforms, which differ according to their level of post-translational modification. The triangle indicates the theoretical MW and pI of the protein.
1D SDS-PAGE of ab83580 before and after treatment with glycosidases to remove oligosaccharides. Lane 1 – MW markers; Lane 2 – ab83580; Lane 3 – ab83580 treated with PNGase F to remove potential N-linked glycans; Lane 4 – ab83580 treated with a glycosidase cocktail to remove potential N- and O-linked glycans. Approximately 5 µg of protein was loaded per lane.Drop in MW after treatment with PNGase F indicates presence of N-linked glycans. Additional bands in lane 3 and lane 4 are glycosidase enzymes.
A sample of ab83580 without carrier protein was reduced and alkylated and focused on a 3-10 IPG strip then run on a 4-20% Tris-HCl 2D gel. Approximately 40 µg of protein was loaded; Gel was stained using coloidal Coomassie Brilliant Blue. Spot train indicates presence of multiple glycoforms of c-Kit.
References for c-Kit protein (Active) (ab83580)
ab83580 has not yet been referenced specifically in any publications.