Anti CDC2 - Cosmo Bio Co.,Ltd.

Antibodies

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Anti CDC2 Antibody

cell division cycle kinase

Catalog No.: AGR-AS06 153
Size: 100UL
Price: ¥112000
$1494
antigen/source: CDC2
host: Rabbit
Purity: Serum
Application: Western Blot
Storage: -20C
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Immunogen: Corn
Reacts with: Arabidopsis thaliana
Barley
Corn
Rice
Vicia fava
Shape: Lyophilized
Molecular Weight: 34-36 (Zea mays)
Purpose: Western Blot: 1: 2000 - 1 : 10 000
Immunolocalization: 1: 500
Optimal dilutions/concentrations should be determined by the end user.



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Seeds of field bean (Vicia faba L. subsp. minor var. Nadwislanski; DANKO Group; Sobiejuchy) were sterilized using sodium hypochlorite (0.3% v/v) and germinated in Petri dishes on wetted filter paper at room temperature. At 4 d after imbibition, dark-grown seedlings with primary roots 25}5 mm long were selected for experiments. During incubations roots were oriented horizontally in a humid chamber and aerated continuously on a rotary water-bath shaker (30 rpm) at 23‹C. Immunocytochemical assays were performed according to the method prescribed earlier (Rybaczek and Maszewski 2006). Excised apical parts of roots (1.5 mm long) were fixed for 45 min (18‹C) in PBS-buffered 3.7% paraformaldehyde, washed several times with PBS and placed in a citric acid-buffered digestion solution (pH 5.0; 37‹C for 45 min) containing 2.5% pectinase (Fluka), 2.5% cellulase (Onozuka R-10; Serva) and 2.5% pectoliase (ICN). After removing the digestion solution, root tips were washed 3 times in PBS, rinsed with distilled water and squashed onto Super Frost Plus glass slides (Menzel-Glaser). Air-dried slides were pretreated with PBS-buffered 5% BSA at 20‹C for 50 min and incubated overnight in a humidified atmosphere (4‹C) with rabbit antibody raised against CDC2 (Agrisera), dissolved in PBS containing 1% BSA (at a dilution of 1:500). Following incubation, slides were washed 3 times with PBS and incubated for 1 h (18‹C) with secondary goat anti-rabbit IgG DyLightR488 antibody (AS09 633, Agrisera; 1:3000). Nuclear DNA was stained with 4f,6-diamidino-2-phenyl-indole (DAPI, 0.4 ƒÊg/ml; Sigma-Aldrich). Following washing with PBS, slides were air dried and embedded in Vectashield Mounting Media for Fluorescence (Vector Laboratories). Observations were made using Optiphot-2 fluorescence microscope (Nikon) equipped with B-2A filter (blue light; ƒÉ 495 nm) for DyLight-conjugated antibodies and UV-2A filter (UV light; ƒÉ 365 nm) for DAPI. All images were recorded at exactly the same time of
 
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