Key features and details
- Rabbit polyclonal to NEK9 (phospho T210)
- Suitable for: WB, ICC/IF, ELISA, IHC-P
- Reacts with: Human
- Isotype: IgG
Product nameAnti-NEK9 (phospho T210) antibody
See all NEK9 primary antibodies
DescriptionRabbit polyclonal to NEK9 (phospho T210)
SpecificityDetects endogenous levels of NEK9 only when phosphorylated at threonine 210.
Tested applicationsSuitable for: WB, ICC/IF, ELISA, IHC-Pmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse
Synthetic phosphopeptide derived from human NEK9 around the phosphorylation site of threonine 210 (A-E-TP-L-V).
- HeLa cells and extracts from HepG2 cells
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride
Without Mg2+ and Ca2+
Concentration information loading...
PurityImmunogen affinity purified
Purification notesThe antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
Our Abpromise guarantee covers the use of ab63553 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500 - 1/1000. Detects a band of approximately 130 kDa (predicted molecular weight: 107 kDa).|
|ICC/IF||1/500 - 1/1000.|
|IHC-P||Use at an assay dependent concentration.|
FunctionPleiotropic regulator of mitotic progression, participating in the control of spindle dynamics and chromosome separation. Phosphorylates different histones, myelin basic protein, beta-casein, and BICD2. Phosphorylates histone H3 on serine and threonine residues and beta-casein on serine residues. Important for G1/S transition and S phase progression.
Tissue specificityMost abundant in heart, liver, kidney and testis. Also expressed in smooth muscle cells and fibroblasts.
Sequence similaritiesBelongs to the protein kinase superfamily. NEK Ser/Thr protein kinase family. NIMA subfamily.
Contains 1 protein kinase domain.
Contains 6 RCC1 repeats.
Developmental stageExpression varied mildly across the cell cycle, with highest expression observed in G1 and stationary-phase cells.
DomainDimerizes through its coiled-coil domain.
modificationsAutophosphorylated on serine and threonine residues. When complexed with FACT, exhibits markedly elevated phosphorylation on Thr-210. During mitosis, not phosphorylated on Thr-210. Phosphorylated by CDK1 in vitro.
Cellular localizationCytoplasm. Nucleus.
- Information by UniProt
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All lanes : Anti-NEK9 (phospho T210) antibody (ab63553) at 1/500 dilution
Lane 1 : extracts from HepG2 cells at 30 µg/ml
Lane 2 : extracts from HepG2 cells at 30 µg with immunizing peptide at 10 µg
Predicted band size: 107 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?
ab63553 at 1/500 dilution staining HeLa cells, without (left) and with (right) immunizing peptide.
Ab63553 staining Human colon. Staining is localised to the cytoplasm and nucleus.
Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffers EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
ab63553 has been referenced in 1 publication.
- Levinsohn JL et al. Somatic Mutations in NEK9 Cause Nevus Comedonicus. Am J Hum Genet 98:1030-1037 (2016). PubMed: 27153399