Product: Phospho-NuMA (Ser395) Antibody
Catalog: AF2373
Description: Rabbit polyclonal antibody to Phospho-NuMA (Ser395)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Dog, Chicken
Mol.Wt.: 238kDa; 238kD(Calculated).
Uniprot: Q14980
RRID: AB_2845387

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, 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:
Dog(83%), Chicken(86%)
Clonality:
Polyclonal
Specificity:
Phospho-NuMA (Ser395) Antibody detects endogenous levels of NuMA only when phosphorylated at Ser395.
RRID:
AB_2845387
Cite Format: Affinity Biosciences Cat# AF2373, RRID:AB_2845387.
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

Centrophilin stabilizes mitotic spindle in mitotic cells; NMP 22; Nuclear matrix protein 22; Nuclear mitotic apparatus protein 1; Nuclear mitotic apparatus protein; NUMA 1; NUMA; NuMA protein; NUMA1; NUMA1_HUMAN; SP H antigen; SP-H antigen; Structural nuclear protein;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Sequence:
MTLHATRGAALLSWVNSLHVADPVEAVLQLQDCSIFIKIIDRIHGTEEGQQILKQPVSERLDFVCSFLQKNRKHPSSPECLVSAQKVLEGSELELAKMTMLLLYHSTMSSKSPRDWEQFEYKIQAELAVILKFVLDHEDGLNLNEDLENFLQKAPVPSTCSSTFPEELSPPSHQAKREIRFLELQKVASSSSGNNFLSGSPASPMGDILQTPQFQMRRLKKQLADERSNRDELELELAENRKLLTEKDAQIAMMQQRIDRLALLNEKQAASPLEPKELEELRDKNESLTMRLHETLKQCQDLKTEKSQMDRKINQLSEENGDLSFKLREFASHLQQLQDALNELTEEHSKATQEWLEKQAQLEKELSAALQDKKCLEEKNEILQGKLSQLEEHLSQLQDNPPQEKGEVLGDVLQLETLKQEAATLAANNTQLQARVEMLETERGQQEAKLLAERGHFEEEKQQLSSLITDLQSSISNLSQAKEELEQASQAHGARLTAQVASLTSELTTLNATIQQQDQELAGLKQQAKEKQAQLAQTLQQQEQASQGLRHQVEQLSSSLKQKEQQLKEVAEKQEATRQDHAQQLATAAEEREASLRERDAALKQLEALEKEKAAKLEILQQQLQVANEARDSAQTSVTQAQREKAELSRKVEELQACVETARQEQHEAQAQVAELELQLRSEQQKATEKERVAQEKDQLQEQLQALKESLKVTKGSLEEEKRRAADALEEQQRCISELKAETRSLVEQHKRERKELEEERAGRKGLEARLQQLGEAHQAETEVLRRELAEAMAAQHTAESECEQLVKEVAAWRERYEDSQQEEAQYGAMFQEQLMTLKEECEKARQELQEAKEKVAGIESHSELQISRQQNELAELHANLARALQQVQEKEVRAQKLADDLSTLQEKMAATSKEVARLETLVRKAGEQQETASRELVKEPARAGDRQPEWLEEQQGRQFCSTQAALQAMEREAEQMGNELERLRAALMESQGQQQEERGQQEREVARLTQERGRAQADLALEKAARAELEMRLQNALNEQRVEFATLQEALAHALTEKEGKDQELAKLRGLEAAQIKELEELRQTVKQLKEQLAKKEKEHASGSGAQSEAAGRTEPTGPKLEALRAEVSKLEQQCQKQQEQADSLERSLEAERASRAERDSALETLQGQLEEKAQELGHSQSALASAQRELAAFRTKVQDHSKAEDEWKAQVARGRQEAERKNSLISSLEEEVSILNRQVLEKEGESKELKRLVMAESEKSQKLEERLRLLQAETASNSARAAERSSALREEVQSLREEAEKQRVASENLRQELTSQAERAEELGQELKAWQEKFFQKEQALSTLQLEHTSTQALVSELLPAKHLCQQLQAEQAAAEKRHREELEQSKQAAGGLRAELLRAQRELGELIPLRQKVAEQERTAQQLRAEKASYAEQLSMLKKAHGLLAEENRGLGERANLGRQFLEVELDQAREKYVQELAAVRADAETRLAEVQREAQSTARELEVMTAKYEGAKVKVLEERQRFQEERQKLTAQVEQLEVFQREQTKQVEELSKKLADSDQASKVQQQKLKAVQAQGGESQQEAQRLQAQLNELQAQLSQKEQAAEHYKLQMEKAKTHYDAKKQQNQELQEQLRSLEQLQKENKELRAEAERLGHELQQAGLKTKEAEQTCRHLTAQVRSLEAQVAHADQQLRDLGKFQVATDALKSREPQAKPQLDLSIDSLDLSCEEGTPLSITSKLPRTQPDGTSVPGEPASPISQRLPPKVESLESLYFTPIPARSQAPLESSLDSLGDVFLDSGRKTRSARRRTTQIINITMTKKLDVEEPDSANSSFYSTRSAPASQASLRATSSTQSLARLGSPDYGNSALLSLPGYRPTTRSSARRSQAGVSSGAPPGRNSFYMGTCQDEPEQLDDWNRIAELQQRNRVCPPHLKTCYPLESRPSLSLGTITDEEMKTGDPQETLRRASMQPIQIAEGTGITTRQQRKRVSLEPHQGPGTPESKKATSCFPRPMTPRDRHEGRKQSTTEAQKKAAPASTKQADRRQSMAFSILNTPKKLGNSLLRRGASKKALSKASPNTRSGTRRSPRIATTTASAATAAAIGATPRAKGKAKH

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
Chicken
86
Dog
83
Pig
75
Bovine
75
Sheep
75
Rabbit
73
Horse
67
Xenopus
67
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q14980 As Substrate

Site PTM Type Enzyme
T2 Phosphorylation
T6 Phosphorylation
R42 Methylation
T46 Phosphorylation
K54 Ubiquitination
K73 Ubiquitination
S76 Phosphorylation
S77 Phosphorylation
C80 S-Nitrosylation
S91 Phosphorylation
T107 Phosphorylation
S112 Phosphorylation
K153 Ubiquitination
S161 Phosphorylation
S162 Phosphorylation
T163 Phosphorylation
S169 Phosphorylation
S172 Phosphorylation
K176 Ubiquitination
S198 Phosphorylation
S200 Phosphorylation
S203 Phosphorylation
T211 Phosphorylation
K247 Ubiquitination
K267 Ubiquitination
S271 Phosphorylation
K284 Ubiquitination
K297 Ubiquitination
K303 Acetylation
R311 Methylation
K312 Ubiquitination
K326 Ubiquitination
K358 Ubiquitination
K364 Ubiquitination
K379 Acetylation
K379 Ubiquitination
K386 Ubiquitination
S388 Phosphorylation
S395 Phosphorylation
K405 Ubiquitination
K419 Ubiquitination
T441 Phosphorylation
K449 Ubiquitination
K531 Ubiquitination
K561 Ubiquitination
T587 Phosphorylation
S595 Phosphorylation
K604 Ubiquitination
K616 Ubiquitination
S633 Phosphorylation
T636 Phosphorylation
S637 Phosphorylation
K697 Ubiquitination
K708 Ubiquitination
K740 Ubiquitination
K755 Ubiquitination
S801 Phosphorylation
S820 Phosphorylation
K855 Ubiquitination
K891 Acetylation
K891 Ubiquitination
K897 Ubiquitination
K908 Ubiquitination
K914 Ubiquitination
S934 Phosphorylation
K939 Ubiquitination
T963 Phosphorylation
S991 Phosphorylation
K1024 Ubiquitination
T1047 Phosphorylation
K1059 Ubiquitination
K1068 Ubiquitination
K1078 Ubiquitination
S1103 Phosphorylation
S1105 Phosphorylation
S1109 Phosphorylation
K1121 Acetylation
K1121 Ubiquitination
K1131 Ubiquitination
K1138 Ubiquitination
S1145 Phosphorylation
S1149 Phosphorylation
K1174 Ubiquitination
S1187 Phosphorylation
S1203 Phosphorylation
K1210 Ubiquitination
K1223 Ubiquitination
S1225 Phosphorylation
S1228 Phosphorylation
S1229 Phosphorylation
S1262 Phosphorylation
S1278 Phosphorylation
S1280 Phosphorylation
K1330 Ubiquitination
K1335 Ubiquitination
K1339 Ubiquitination
K1364 Ubiquitination
C1367 S-Nitrosylation
K1379 Ubiquitination
K1389 Ubiquitination
K1430 Ubiquitination
S1438 Phosphorylation
K1441 Ubiquitination
K1442 Ubiquitination
R1452 Methylation
K1475 Ubiquitination
T1489 Phosphorylation
T1509 Phosphorylation
K1511 Acetylation
K1511 Ubiquitination
K1518 Ubiquitination
K1573 Ubiquitination
S1582 Phosphorylation
S1601 Phosphorylation
K1603 Ubiquitination
K1624 Acetylation
K1625 Ubiquitination
K1643 Ubiquitination
K1665 Ubiquitination
K1699 Ubiquitination
T1704 Phosphorylation
K1708 Ubiquitination
K1715 Ubiquitination
S1721 Phosphorylation
S1724 Phosphorylation
S1728 Phosphorylation
T1733 Phosphorylation
S1739 Phosphorylation
K1740 Ubiquitination
T1744 Phosphorylation
T1749 Phosphorylation
S1750 Phosphorylation
S1757 Phosphorylation
S1760 Phosphorylation
K1766 Acetylation
K1766 Sumoylation
K1766 Ubiquitination
S1769 Phosphorylation
S1772 Phosphorylation
Y1774 Phosphorylation P00519 (ABL1)
T1776 Phosphorylation P24941 (CDK2)
S1782 Phosphorylation
S1788 Phosphorylation
S1789 Phosphorylation
S1792 Phosphorylation
S1800 Phosphorylation
R1802 Methylation
T1804 Phosphorylation O14965 (AURKA)
S1806 Phosphorylation
T1811 Phosphorylation Q96GD4 (AURKB) , O14965 (AURKA)
T1812 Phosphorylation O14965 (AURKA)
T1818 Phosphorylation
T1820 Phosphorylation
K1822 Ubiquitination
S1830 Phosphorylation
S1833 Phosphorylation
S1834 Phosphorylation
Y1836 Phosphorylation
S1837 Phosphorylation
T1838 Phosphorylation
S1840 Phosphorylation
S1844 O-Glycosylation
S1844 Phosphorylation
S1847 Phosphorylation
R1849 Methylation
S1852 Phosphorylation
S1853 Phosphorylation
T1854 Phosphorylation
R1859 Methylation
S1862 Phosphorylation
Y1865 Phosphorylation
S1868 Phosphorylation
S1872 Phosphorylation
Y1876 Phosphorylation
T1879 Phosphorylation
T1880 Phosphorylation
S1882 Phosphorylation
S1883 Phosphorylation
S1887 Phosphorylation O14965 (AURKA)
S1892 Phosphorylation
S1901 Phosphorylation
Y1903 Phosphorylation
K1935 Ubiquitination
T1936 Phosphorylation
S1942 Phosphorylation
S1945 Phosphorylation
S1947 Phosphorylation
T1950 Phosphorylation
T1952 Phosphorylation
K1957 Ubiquitination
T1964 Phosphorylation
S1969 Phosphorylation O14965 (AURKA) , Q96GD4 (AURKB)
K1988 Acetylation
S1991 Phosphorylation O14965 (AURKA)
T2000 Phosphorylation
S2003 Phosphorylation
K2004 Acetylation
K2004 Ubiquitination
T2015 Phosphorylation P06493 (CDK1)
S2026 Phosphorylation
T2027 Phosphorylation
T2028 Phosphorylation
S2047 Phosphorylation Q96GD4 (AURKB) , O14965 (AURKA)
S2051 Phosphorylation
T2055 Phosphorylation P06493 (CDK1) , Q96GD4 (AURKB)
K2057 Acetylation
S2062 Phosphorylation
K2070 Acetylation
K2071 Acetylation
S2074 Phosphorylation
S2077 Phosphorylation
T2080 Phosphorylation
T2084 Phosphorylation O14965 (AURKA)
S2087 Phosphorylation P06493 (CDK1) , O14965 (AURKA)
T2099 Phosphorylation
T2106 Phosphorylation P06493 (CDK1)
K2114 Methylation

Research Backgrounds

Function:

Microtubule (MT)-binding protein that plays a role in the formation and maintenance of the spindle poles and the alignement and the segregation of chromosomes during mitotic cell division. Functions to tether the minus ends of MTs at the spindle poles, which is critical for the establishment and maintenance of the spindle poles. Plays a role in the establishment of the mitotic spindle orientation during metaphase and elongation during anaphase in a dynein-dynactin-dependent manner. In metaphase, part of a ternary complex composed of GPSM2 and G(i) alpha proteins, that regulates the recruitment and anchorage of the dynein-dynactin complex in the mitotic cell cortex regions situated above the two spindle poles, and hence regulates the correct oritentation of the mitotic spindle. During anaphase, mediates the recruitment and accumulation of the dynein-dynactin complex at the cell membrane of the polar cortical region through direct association with phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2), and hence participates in the regulation of the spindle elongation and chromosome segregation. Binds also to other polyanionic phosphoinositides, such as phosphatidylinositol 3-phosphate (PIP), lysophosphatidic acid (LPA) and phosphatidylinositol triphosphate (PIP3), in vitro. Also required for proper orientation of the mitotic spindle during asymmetric cell divisions. Plays a role in mitotic MT aster assembly. Involved in anastral spindle assembly. Positively regulates TNKS protein localization to spindle poles in mitosis. Highly abundant component of the nuclear matrix where it may serve a non-mitotic structural role, occupies the majority of the nuclear volume. Required for epidermal differentiation and hair follicle morphogenesis (By similarity).

PTMs:

Phosphorylation and dephosphorylation on Thr-2055 regulates the extent of cortical NUMA1 and the dynein-dynactin complex localization during mitotic metaphase and anaphase. In metaphase, phosphorylation on Thr-2055 occurs in a kinase CDK1-dependent manner; this phosphorylation maintains low levels of cortical dynein-dynactin complex at metaphase, and hence proper spindle positioning. In anaphase, dephosphorylated on Thr-2055 by phosphatase PPP2CA; this dephosphorylation stimulates its membrane association and with the dynein-dynactin complex its enrichment at the cell cortex, and hence robust spindle elongation. Probably also phosphorylated on Thr-2015 and Ser-2087 by CDK1; these phosphorylations may regulate its cell cortex recruitment during metaphase and anaphase. Phosphorylated on Thr-1047, Ser-1769, Ser-1772, Ser-1789 and Ser-1834 by PLK1; these phosphorylations induce cortical dynein-dynactin complex dissociation from the NUMA1-GPSM2 complex and negatively regulates cortical dynein-dynactin complex localization.

ADP-ribosylated by TNKS at the onset of mitosis; ADP-ribosylation is not required for its localization to spindle poles.

O-glycosylated during cytokinesis at sites identical or close to phosphorylation sites, this interferes with the phosphorylation status.

Ubiquitinated with 'Lys-63'-linked polyubiquitin chains. Deubiquitination by the BRISC complex is important for the incorporation of NUMA1 into mitotic spindle poles and normal spindle pole function, probably by modulating interactions between NUMA1, dynein-dynactin complex and importin-beta.

Subcellular Location:

Nucleus. Nucleus>Nucleoplasm. Nucleus matrix. Chromosome. Cytoplasm>Cytoskeleton. Cytoplasm>Cytoskeleton>Microtubule organizing center>Centrosome. Cytoplasm>Cytoskeleton>Spindle pole. Cytoplasm>Cell cortex. Cell membrane>Lipid-anchor>Cytoplasmic side. Lateral cell membrane.
Note: Mitotic cell cycle-dependent shuttling protein that relocalizes from the interphase nucleus to the spindle poles and cell cortex (PubMed:1541636, PubMed:10811826). The localization to the spindle poles is regulated by AAAS (PubMed:26246606). In interphase, resides in the nuclear matrix (PubMed:1541630, PubMed:1541636, PubMed:23921553). In prophase, restricted to the interchromatin or condensed chromosome space (PubMed:10811826). In prometaphase, after nuclear envelope disassembly, forms aggregates both in the spindle midzone and at duplicated centrosomes and astral microtubules (MTs) of the bipolar spindle apparatus (PubMed:10811826). Translocates from the spindle midzone towards the spindle poles along spindle fibers in a MT- and dynein-dynactin-dependent manner until the anaphase onset (PubMed:1541636, PubMed:10811826). In metaphase, recruited to the polar cortical region in a GPSM2- and GNAI1-dependent manner (PubMed:23870127, PubMed:24109598, PubMed:24996901). Excluded from the metaphase equatorial cortical region in a RanGTP-dependent manner (PubMed:22327364, PubMed:23870127). Phosphorylation on Thr-2055 by CDK1 results in its localization at spindle poles in metaphase, but not at the cell cortex (PubMed:23921553). In anaphase, recruited and anchored at the cell membrane of the polar cortical region in a EPB41-, EPB41L2-, phosphatidylinositol-dependent and GPSM2- and G(i) alpha proteins-independent manner (PubMed:23870127, PubMed:24996901, PubMed:24109598, PubMed:24371089). Excluded from the anaphase equatorial region of the cell cortex in a RACGAP1- and KIF23-dependent and RanGTP-independent manner (PubMed:24996901). Associated with astral MTs emanating from the spindle poles during anaphase (PubMed:12445386, PubMed:24996901). Nonphosphorylated Thr-2055 localizes at the cell cortex, weakly during metaphase and more prominently during anaphase in a phosphatase PPP2CA-dependent manner (PubMed:23921553). As mitosis progresses it reassociates with telophase chromosomes very early during nuclear reformation, before substantial accumulation of lamins on chromosomal surfaces is evident (PubMed:1541636). Localizes to the tips of cortical MTs in prometaphase (PubMed:26765568). Localizes along MTs and specifically to both MT plus and minus ends (PubMed:26765568). Accumulates also at MT tips near the cell periphery (PubMed:26765568). Colocalizes with GPSM2 at mitotic spindle poles during mitosis (PubMed:11781568, PubMed:21816348). Colocalizes with SPAG5 at mitotic spindle at prometaphase and at mitotic spindle poles at metaphase and anaphase (PubMed:27462074). Colocalizes with ABRO1 at mitotic spindle poles (PubMed:26195665). Colocalized with TNKS from prophase through to anaphase in mitosis (PubMed:16076287). Colocalizes with tubulin alpha (PubMed:12445386). CCSAP is essential for its centrosomal localization (PubMed:26562023). In horizontally retinal progenitor dividing cells, localized to the lateral cortical region (By similarity).

Cytoplasm>Cytosol. Cytoplasm>Cytoskeleton>Microtubule organizing center>Centrosome. Cytoplasm>Cytoskeleton>Spindle pole.
Note: During interphase, mainly clustered at the centrosomal region in the cytosol. After entry into mitosis, detected at mitotic spindle poles.

Cytoplasm>Cytosol. Cytoplasm>Cytoskeleton>Microtubule organizing center>Centrosome. Cytoplasm>Cytoskeleton>Spindle pole.
Note: During interphase, mainly clustered at the centrosomal region in the cytosol. After entry into mitosis, detected at mitotic spindle poles.

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

Homodimer. Also forms multiarm oligomers by association of C-terminal tail domains, oligomers may further assemble to form a hexagonal nuclear lattice-like network. Associates with the dynein-dynactin complex; this association promotes the transport and accumulation of NUMA1 at the mitotic spindle poles that is inhibited by the BRISC complex in a PLK1-dependent manner. Part of a spindle orientation complex at least composed of GNAI1, GPSM2 and NUMA1. Interacts (via C-terminus) with microtubules (MTs); this interaction is direct and promotes both MT bundle formation and stability in a dynein-dynactin complex- and CDK1-independent manner. Interacts with EPB41 and EPB41L2; these interactions are negatively regulated by CDK1 during metaphase and are important for anaphase-specific localization of NUMA1 in symmetrically dividing cells. Interacts (via C-terminus) with GPSM2 (via TPR repeats); this interaction is direct, prevented by competitive binding of INSC, is inhibited in a PLK1-dependent manner, blocks the association of NUMA1 with MTs and inhibits NUMA1-induced MT bundle formation, prevents the association of NUMA1 with SPAG5, induces mitotic spindle pole localization of GPSM2, both metaphase cell cortex localization of NUMA1 and mitotic spindle organization. Does not interact with GPSM2 during anaphase. Interacts (via C-terminus) with the nuclear importin alpha/importin beta receptor; this interaction is inhibited by RanGTP. Interacts (via C-terminus) with KPNB1; this interaction is inhibited by RanGTP and the BRISC complex. Interacts with ABRAXAS2 and the BRISC complex; these interactions regulate mitotic spindle assembly. Interacts (via N-terminal end of the coiled-coil domain) with RAE1; this interaction promotes mitotic spindle formation. Interacts (via C-terminus) with SPAG5 (via C-terminus); this interaction promotes the recruitment of SPAG5 to the MTs at spindle poles in a dynein-dynactin-dependent manner and regulates mitotic spindle organization and proper chromosome alignment during mitosis. Interacts with TNKS; this interaction occurs at the onset of mitosis. Interacts with TNKS2. Interacts with tubulin.

Family&Domains:

The C-terminal tubulin-binding domain mediates direct binding to microtubules, independently of dynein-dynactin complex, and induces their bundling and stabilization (PubMed:11956313). The 4.1-binding domain is necessary for its cortical stability and spindle orientation (PubMed:24109598).

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