Product: Phospho-c-Kit (Tyr721) Antibody
Catalog: AF3153
Description: Rabbit polyclonal antibody to Phospho-c-Kit (Tyr721)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Rabbit, Dog
Mol.Wt.: 120,145kDa; 110kD(Calculated).
Uniprot: P10721
RRID: AB_2834588

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:100-1:500, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Rabbit(100%), Dog(100%)
Clonality:
Polyclonal
Specificity:
Phospho-c-Kit (Tyr721) Antibody detects endogenous levels of c-Kit only when phosphorylated at Tyrosine 721.
RRID:
AB_2834588
Cite Format: Affinity Biosciences Cat# AF3153, RRID:AB_2834588.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

C Kit; c-Kit; c-Kit Ligand; CD117; Kit; Kit Ligand; KIT oncogene; KIT proto oncogene receptor tyrosine kinase; KIT_HUMAN; Mast cell growth factor receptor; Mast/stem cell growth factor receptor Kit; MGF; p145 c-kit; PBT; Piebald trait protein; Proto oncogene c Kit; Proto oncogene tyrosine protein kinase Kit; Proto-oncogene c-Kit; SCF Receptor; SCFR; soluble KIT variant 1; Steel Factor Receptor; Stem cell factor receptor; tyrosine protein kinase Kit; Tyrosine-protein kinase Kit; v kit Hardy Zuckerman 4 feline sarcoma viral oncogene homolog; v kit Hardy Zuckerman 4 feline sarcoma viral oncogene like protein; v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P10721 KIT_HUMAN:

Isoform 1 and isoform 2 are detected in spermatogonia and Leydig cells. Isoform 3 is detected in round spermatids, elongating spermatids and spermatozoa (at protein level). Widely expressed. Detected in the hematopoietic system, the gastrointestinal system, in melanocytes and in germ cells.

Description:
KIT encodes the human homolog of the proto-oncogene c-kit. C-kit was first identified as the cellular homolog of the feline sarcoma viral oncogene v-kit. KIT is a type 3 transmembrane receptor for MGF (mast cell growth factor, also known as stem cell factor). Mutations in KIT are associated with gastrointestinal stromal tumors, mast cell disease, acute myelogenous lukemia, and piebaldism.
Sequence:
MRGARGAWDFLCVLLLLLRVQTGSSQPSVSPGEPSPPSIHPGKSDLIVRVGDEIRLLCTDPGFVKWTFEILDETNENKQNEWITEKAEATNTGKYTCTNKHGLSNSIYVFVRDPAKLFLVDRSLYGKEDNDTLVRCPLTDPEVTNYSLKGCQGKPLPKDLRFIPDPKAGIMIKSVKRAYHRLCLHCSVDQEGKSVLSEKFILKVRPAFKAVPVVSVSKASYLLREGEEFTVTCTIKDVSSSVYSTWKRENSQTKLQEKYNSWHHGDFNYERQATLTISSARVNDSGVFMCYANNTFGSANVTTTLEVVDKGFINIFPMINTTVFVNDGENVDLIVEYEAFPKPEHQQWIYMNRTFTDKWEDYPKSENESNIRYVSELHLTRLKGTEGGTYTFLVSNSDVNAAIAFNVYVNTKPEILTYDRLVNGMLQCVAAGFPEPTIDWYFCPGTEQRCSASVLPVDVQTLNSSGPPFGKLVVQSSIDSSAFKHNGTVECKAYNDVGKTSAYFNFAFKGNNKEQIHPHTLFTPLLIGFVIVAGMMCIIVMILTYKYLQKPMYEVQWKVVEEINGNNYVYIDPTQLPYDHKWEFPRNRLSFGKTLGAGAFGKVVEATAYGLIKSDAAMTVAVKMLKPSAHLTEREALMSELKVLSYLGNHMNIVNLLGACTIGGPTLVITEYCCYGDLLNFLRRKRDSFICSKQEDHAEAALYKNLLHSKESSCSDSTNEYMDMKPGVSYVVPTKADKRRSVRIGSYIERDVTPAIMEDDELALDLEDLLSFSYQVAKGMAFLASKNCIHRDLAARNILLTHGRITKICDFGLARDIKNDSNYVVKGNARLPVKWMAPESIFNCVYTFESDVWSYGIFLWELFSLGSSPYPGMPVDSKFYKMIKEGFRMLSPEHAPAEMYDIMKTCWDADPLKRPTFKQIVQLIEKQISESTNHIYSNLANCSPNRQKPVVDHSVRINSVGSTASSSQPLLVHDDV

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Dog
100
Rabbit
100
Chicken
73
Xenopus
71
Sheep
0
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P10721 As Substrate

Site PTM Type Enzyme
N130 N-Glycosylation
K173 Sumoylation
S215 Phosphorylation
S217 Phosphorylation
T232 Phosphorylation
S240 Phosphorylation
S241 Phosphorylation
N283 N-Glycosylation
N293 N-Glycosylation
N300 N-Glycosylation
N320 N-Glycosylation
N352 N-Glycosylation
T354 Phosphorylation
Y362 Phosphorylation
N367 N-Glycosylation
T488 Phosphorylation
Y494 Phosphorylation
Y547 Phosphorylation P10721 (KIT)
Y553 Phosphorylation P10721 (KIT)
Y568 Phosphorylation P10721 (KIT)
Y570 Phosphorylation P10721 (KIT)
Y578 Phosphorylation
Y609 Phosphorylation
K642 Ubiquitination
Y703 Phosphorylation P10721 (KIT)
T718 Phosphorylation
Y721 Phosphorylation P10721 (KIT)
S729 Phosphorylation
Y730 Phosphorylation P10721 (KIT)
K735 Ubiquitination
K738 Ubiquitination
S741 Phosphorylation P17252 (PRKCA)
S746 Phosphorylation P17252 (PRKCA)
Y747 Phosphorylation
T801 Phosphorylation
S821 Phosphorylation
Y823 Phosphorylation P10721 (KIT)
S891 Phosphorylation
Y900 Phosphorylation P12931 (SRC) , P10721 (KIT)
S931 Phosphorylation
Y936 Phosphorylation P10721 (KIT)
S937 Phosphorylation
S943 Phosphorylation
S959 Phosphorylation

PTMs - P10721 As Enzyme

Substrate Site Source
P10721 (KIT) Y547 Uniprot
P10721 (KIT) Y553 Uniprot
P10721 (KIT) Y568 Uniprot
P10721 (KIT) Y570 Uniprot
P10721 (KIT) Y703 Uniprot
P10721 (KIT) Y721 Uniprot
P10721 (KIT) Y730 Uniprot
P10721 (KIT) Y823 Uniprot
P10721 (KIT) Y900 Uniprot
P10721 (KIT) Y936 Uniprot
P42224-1 (STAT1) Y701 Uniprot
Q9GZY6 (LAT2) Y110 Uniprot
Q9GZY6 (LAT2) Y118 Uniprot

Research Backgrounds

Function:

Tyrosine-protein kinase that acts as cell-surface receptor for the cytokine KITLG/SCF and plays an essential role in the regulation of cell survival and proliferation, hematopoiesis, stem cell maintenance, gametogenesis, mast cell development, migration and function, and in melanogenesis. In response to KITLG/SCF binding, KIT can activate several signaling pathways. Phosphorylates PIK3R1, PLCG1, SH2B2/APS and CBL. Activates the AKT1 signaling pathway by phosphorylation of PIK3R1, the regulatory subunit of phosphatidylinositol 3-kinase. Activated KIT also transmits signals via GRB2 and activation of RAS, RAF1 and the MAP kinases MAPK1/ERK2 and/or MAPK3/ERK1. Promotes activation of STAT family members STAT1, STAT3, STAT5A and STAT5B. Activation of PLCG1 leads to the production of the cellular signaling molecules diacylglycerol and inositol 1,4,5-trisphosphate. KIT signaling is modulated by protein phosphatases, and by rapid internalization and degradation of the receptor. Activated KIT promotes phosphorylation of the protein phosphatases PTPN6/SHP-1 and PTPRU, and of the transcription factors STAT1, STAT3, STAT5A and STAT5B. Promotes phosphorylation of PIK3R1, CBL, CRK (isoform Crk-II), LYN, MAPK1/ERK2 and/or MAPK3/ERK1, PLCG1, SRC and SHC1.

PTMs:

Ubiquitinated 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.

Subcellular Location:

Cell membrane>Single-pass type I membrane protein.

Cell membrane>Single-pass type I membrane protein.

Cytoplasm.
Note: Detected in the cytoplasm of spermatozoa, especially in the equatorial and subacrosomal region of the sperm head.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Isoform 1 and isoform 2 are detected in spermatogonia and Leydig cells. Isoform 3 is detected in round spermatids, elongating spermatids and spermatozoa (at protein level). Widely expressed. Detected in the hematopoietic system, the gastrointestinal system, in melanocytes and in germ cells.

Subunit Structure:

Monomer in the absence of bound KITLG/SCF. Homodimer in the presence of bound KITLG/SCF, forming a heterotetramer with two KITLG/SCF molecules. Interacts (via phosphorylated tyrosine residues) with the adapter proteins GRB2 and GRB7 (via SH2 domain), and SH2B2/APS. Interacts (via C-terminus) with MPDZ (via the tenth PDZ domain). Interacts (via phosphorylated tyrosine residues) with PIK3R1 and PIK3 catalytic subunit. Interacts (via phosphorylated tyrosine) with CRK (isoform Crk-II), FYN, SHC1 and MATK/CHK (via SH2 domain). Interacts with LYN and FES/FPS. Interacts (via phosphorylated tyrosine residues) with the protein phosphatases PTPN6/SHP-1 (via SH2 domain), PTPN11/SHP-2 (via SH2 domain) and PTPRU. Interacts with PLCG1. Interacts with DOK1 and TEC. Interacts (KITLG/SCF-bound) with IL1RL1. Interacts with IL1RAP (independent of stimulation with KITLG/SCF). A mast cell-specific KITLG/SCF-induced interleukin-33 signaling complex contains IL1RL1, IL1RAP, KIT and MYD88.

Family&Domains:

Belongs to the protein kinase superfamily. Tyr protein kinase family. CSF-1/PDGF receptor subfamily.

Research Fields

· Environmental Information Processing > Signal transduction > MAPK signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Ras signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Rap1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signaling molecules and interaction > Cytokine-cytokine receptor interaction.   (View pathway)

· Environmental Information Processing > Signal transduction > Phospholipase D signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Acute myeloid leukemia.   (View pathway)

· Human Diseases > Cancers: Specific types > Breast cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Central carbon metabolism in cancer.   (View pathway)

· Organismal Systems > Immune system > Hematopoietic cell lineage.   (View pathway)

· Organismal Systems > Endocrine system > Melanogenesis.

References

1). Cytological and spectroscopic characteristics of c‐KIT N822K mutation in core binding factor acute myeloid leukemia cells. Journal of Biophotonics (PubMed: 32390317) [IF=2.8]

Application: WB    Species: human    Sample: AML cells

Figure 6. |The effects of genetic abnormality-induced cytological and spectral alterations in AML cells. (a) Ratio of clone formation. (b) Relative contents of Raman bands screened by CRT algorithm. (c) Western blot analysis showed the c-KIT and p-c-KIT expression, GAPDH served as loading control, along with (d) ratio of phosphorylated c-KIT to total c-KIT. *P<0.05.

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