Phospho-CENPA (Ser7) Antibody - #AF2330

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Product: Phospho-CENPA (Ser7) Antibody
Catalog: AF2330
Description: Rabbit polyclonal antibody to Phospho-CENPA (Ser7)
Application: WB IHC
Cited expt.: WB
Reactivity: Human, Mouse, Rat
Prediction: Pig, Horse, Rabbit
Mol.Wt.: 17kDa; 16kD(Calculated).
Uniprot: P49450
RRID: AB_2845344

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MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user. For optimal experimental results, antibody reuse is not recommended.
*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(82%), Horse(90%), Rabbit(91%)
Clonality:
Polyclonal
Specificity:
Phospho-CENPA (Ser7) Antibody detects endogenous levels of CENPA only when phosphorylated at Ser7.
RRID:
AB_2845344
Cite Format: Affinity Biosciences Cat# AF2330, RRID:AB_2845344.
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

CENP A; CENP-A; cenpa; CENPA_HUMAN; Centromere autoantigen A; Centromere protein A 17kDa; Centromere protein A; Histone H3 like centromeric protein A; Histone H3-like centromeric protein A;

Immunogens

Immunogen:

A synthesized peptide derived from human CENPA around the phosphorylation site of Ser7.

Uniprot:

P49450(CENPA_HUMAN) >>Visit HPA database.

Gene(ID):
CENPA(1058)
Sequence:
MGPRRRSRKPEAPRRRSPSPTPTPGPSRRGPSLGASSHQHSRRRQGWLKEIRKLQKSTHLLIRKLPFSRLAREICVKFTRGVDFNWQAQALLALQEAAEAFLVHLFEDAYLLTLHAGRVTLFPKDVQLARRIRGLEEGLG

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
Rabbit
91
Horse
90
Pig
82
Bovine
73
Sheep
0
Dog
0
Xenopus
0
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

Research Backgrounds

Function:

Histone H3-like nucleosomal protein that is specifically found in centromeric nucleosomes. Replaces conventional H3 in the nucleosome core of centromeric chromatin at the inner plate of the kinetochore. The presence of CENPA subtly modifies the nucleosome structure and the way DNA is wrapped around the nucleosome and gives rise to protruding DNA ends that are less well-ordered and rigid compared to nucleosomes containing histone H3. May serve as an epigenetic mark that propagates centromere identity through replication and cell division. Required for recruitment and assembly of kinetochore proteins, and as a consequence required for progress through mitosis, chromosome segregation and cytokinesis.

PTMs:

Ubiquitinated (Probable). Interaction with herpes virus HSV-1 ICP0 protein, leads to its degradation by the proteasome pathway.

Trimethylated by NTMT1 at the N-terminal glycine after cleavage of Met-1. Methylation is low before incorporation into nucleosomes and increases with cell cycle progression, with the highest levels in mitotic nucleosomes.

Phosphorylated by CDK1 at Ser-68 during early mitosis; this abolishes association with chromatin and centromeres, prevents interaction with HJURP and thereby prevents premature assembly of CENPA into centromeres. Dephosphorylated at Ser-68 by PPP1CA during late mitosis. Phosphorylation of Ser-7 by AURKA and AURKB during prophase is required for localization of AURKA and AURKB at inner centromere and is essential for normal cytokinesis. Initial phosphorylation during prophase is mediated by AURKA and is maintained by AURKB.

Poly-ADP-ribosylated by PARP1.

Subcellular Location:

Nucleus. Chromosome>Centromere>Kinetochore. Chromosome>Centromere.
Note: Localizes exclusively in the kinetochore domain of centromeres. Occupies a compact domain at the inner kinetochore plate stretching across 2 thirds of the length of the constriction but encompassing only one third of the constriction width and height (PubMed:19114591). Phosphorylation at Ser-68 during early mitosis abolishes association with chromatin and centromeres and results in dispersed nuclear location (PubMed:25556658).

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

The CATD (CENPA targeting domain) region is responsible for the more compact structure of nucleosomes containing CENPA (PubMed:15282608). It is necessary and sufficient to mediate the localization into centromeres (PubMed:7962047, PubMed:15282608).

Belongs to the histone H3 family.

References

1). A Tumor Suppressive Role of CYLD as a Novel Potential DUB of Aurora B in Cervical Cancer. Clinical Medicine Insights. Oncology, 2023 (PubMed: 37359274) [IF=2.2]

Application: WB    Species: human    Sample:

Figure 3. CYLD regulates Aurora B activity and function. (A) HeLa cells transfected with CYLD or control plasmid were synchronized by nocodazole for 18 h before harvested. Mitotic HeLa cells were collected by mitotic shake-off. The collected cells were then continued to incubate for the indicated time points (0, 1, 3 h) and following immunoblotted with indicated antibodies. (B) HeLa cells transfected with control or Flag-CYLD plasmid were synchronized by nocodazole for 18 h and then were fixed for IF analysis with indicated antibodies. Scale bar represents 2 μm. (C) Luciferase control and 2 clones of CYLD knock-down stable HeLa cells were generated and harvested for WB assay with indicated antibodies. (D) Luciferase control or CYLD knock-down stable HeLa cells generated above were synchronized by nocodazole for 18 h and then fixed for IF analysis with indicated antibodies. Scale bar represents 2 μm. (E) HeLa cells expressing H2B-GFP were transfected with control or CYLD plasmid; 36 h later, cells were imaged at every 5 min. Around 100 cells in each group were randomly chosen for statistics. The upper panel showed the representative fluorescence video microscopy series from the onset of mitosis to monitor chromosome dynamics and the lower panel displayed the statistical results. Cell numbers were 123 (Control) and 123 (CYLD). P value was calculated using unpaired T-test. Error bars represent the standard deviation. Scale bar represents 5 μm. (F) Control and CYLD stable knock-down SiHa cervical cancer cells were plated and cultured for 24 h; cells were then stained with Hoechst. Later, more than 30 random pictures of every group were captured under a confocal microscope. Finally, we counted and summed up the proportion of cells with micronuclei. The left panel showed a representative micronuclei microscopy of the cells and the right panel displayed the statistical results. For each group, the cell number is more than 1000. P value was calculated using unpaired T-test. Scale bar represents 2 μm.

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