Rabbit anti-CEP97 Antibody, Affinity Purified A301-945A from Bethyl Laboratories, Inc.

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Nature communications
January 25 2019

Fig 1: PACSIN1 functions at the CV stage prior to CP110 loss. a Quantification of CP110 dots in serum starved RPE-1 cells treated with siRNAs for 72 h, serum starved for the last 24 h, and stained with CP110, Actub and CEP164 antibodies. CP110 localization on both centrioles (2 dots) or only the daughter centriole (1 dot) was quantified (left panel, siCtrl = 100, siPACS1 = 63 cells). b Quantification of CEP97 dots in cells treated as in a and stained with CEP97, Actub, and CEP164 antibodies. CEP97 localization on the centrioles was quantified as in a(siCtrl = 201, siPACS1 = 125 cells). c Quantification of SMOM2-GFP centrosomal accumulation in cells treated as in a, and stained with Actub and CEP164 antibodies. “Preciliary” indicates SMOM2-GFP vesicle accumulation at the MC without axonemes (siCtrl = 100, siPACS1 = 67 cells). Scale bar: 2 μm for images in a–c. d Quantification of MC with non-ciliary distal appendages structures from cells treated as in a. Pooled data from 3 independent experiments (siCtrl = 56, siPACS1 = 41 cells). Representative electron micrographs of quantified MCs shown on right. Scale bar: 200 nm. MC mother centriole, DAV distal appendages vesicles, CV ciliary vesicle, or DA non-membrane-associated distal appendages. e–g Quantification of TZ accumulation of TMEM67 (e, siCtrl = 113, siPACS1 = 89 cells), B9D2-GFP (f, siCtrl = 365, siPACS1 = 250 cells), RPGRIP1L (g, siCtrl = 70, siPACS1 = 63 cells) in cells treated as in a. h mIFT20-GFP cells treated as in a, stained with Actub and CEP164 antibodies, and quantified for mIFT20 fluorescence intensity at the MC/BB (siCtrl = 46, siPACS1 = 43 cells). Scale bar: 2 μm for images in e–h. In a–c, and e–h, representative images of quantified cells shown on right and means ± SEM, n = 3 independent experiments. Two-tailed t-test analysis as compared with siCtrl was performed. **P < 0.001, ***P < 0.0001
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Nature communications
February 14 2020

Fig 2: CEP44 is a centriolar protein necessary for the CCC mechanism.a IF of cycling RPE1 cells showing that CEP44 binds to the new dCs during G2. While S phase cells were detected by EdU stain, G1 and G2 cells were discerned by the lack of EdU stain and the number of centrin1 signals (centrioles). b IF of cells after 72 h of depletion. In the siCEP44 sample (lower panel) G1 cells contained less centrosomes as judged by the number of γ-tubulin foci (Cenp-F in Supplementary Fig. 1b). c Quantification of b. 80.4 ± 5.0% of G1 cells contained <2 centrosomes. d IF of 60 h siRNA treated RPE1 cells in G1. While the G1 control cells contained 2 defined PCM foci (γ-tubulin) accompanied by equal number of CEP44 foci, in the siCEP44 sample the loss of CEP44 correlated with inefficient PCM (γ-tubulin) recruitment to only one (bottom panel) or both centrosomes (middle panel) (Cenp-F, Supplementary Fig. 1g). e Quantification of CEP44 loss in d. 65.1 ± 7.3% of G1 cells contained <2 CEP44 foci. f Quantification of γ-tubulin defined foci in d. 60.7 ± 4.2% of G1 cells contained <2 defined γ-tubulin signals. g, h G2 CEP44-depleted cells after 60 h of CEP44 depletion showed a mild centriole duplication defect as judged by the counting of CEP97 foci (<4). g Quantification of h. 30.3 ± 4.1% of CEP44-depleted cells contained <4 centrioles. i MT regrowth assay in RPE1 C-Nap1 KO cells. The non-converted daughter centrosome without CEP164 staining regrew lower numbers of MTs (5.1 ± 2.7 MTs/centrosome) than the siControl daughter centrosomes (10.3 ± 3.0 MTs/centrosome) upon cold treatment and MT regrowth. j Quantification of i. k Loss of CEP44 leads to misalignment of mitotic spindles (>65%) generating either bipolar asymmetric spindles (51.3 ± 4.7%) or mono-/multipolar ones (14.3 ± 4.0). l Quantification of k. (a, b, d, h, i, k, scale bars: 10 μm, magnification scale bars: 1 μm; c, e, f, g, j and l data are presented as mean ± s.d., all statistics were derived from two-tail unpaired t-test analysis of n = 6 biologically independent experiments and source data are provided as a Source Data file).

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Rabbit anti-CEP97 Antibody, Affinity Purified

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The Rabbit anti-CEP97 Antibody, Affinity Purified from Bethyl Laboratories, Inc. is a Rabbit Polyclonal antibody to CEP97. This antibody recognizes Homo Sapiens, and Human antigen. The Rabbit anti-CEP97 Antibody, Affinity Purified has been shown to work in the following applications: Precipitation, Immunoprecipitation, and Western Blot.

Description

The epitope recognized by A301-945A maps to a region between residue 625 and 675 of human centrosomal protein 97kDa using the numbering given in entry NP_078824.2 (GeneID 79598)

Nature communications

Nature communications

Rabbit anti-CEP97 Antibody, Affinity Purified from Bethyl Laboratories, Inc.

Description

Bethyl Laboratories, Inc.

Citations (6)

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Rabbit anti-CEP97 Antibody, Affinity Purified from Bethyl Laboratories, Inc.

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