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CDC2 | Cell-division-cycle kinase 2

AS06 153  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: A. thaliana, H. vulgare, O. sativa, P. yunnaniensis, Z. mays, V.faba

CDC2 | Cell-division-cycle kinase 2 in the group Antibodies Plant/Algal  / DNA/RNA/Cell Cycle / Cell cycle  at Agrisera AB (Antibodies for research) (AS06 153)
CDC2 | Cell-division-cycle kinase 2



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Product Information

Immunogen

KLH-conjugated synthetic peptide derived form Zea mays CDC2 sequence P23111, well conserved in other plant species

Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 µl
Reconstitution For reconstitution add 50 µl of sterile water
Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Tested applications Immunolocalization (IL), Western blot (WB)
Recommended dilution 1 : 500 (IL), 1 : 2000-1 : 10 000 (WB)
Expected | apparent MW

34-36 (Zea mays)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Hordeum vulgare,Vicia faba, Oryza sativa, Pinus yunnanensi, Zea mays
Predicted reactivity Pisum sativum, Solanum lycopersicum, Triticum aestivum, Pinus contorta, Populus balsamifera, Physcomitrium patens
Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples Application example

CDC2 immunolocalization in plant tissue

Seeds of field bean (Vicia faba L. subsp. minor var. Nadwiślański; 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-Gläser). 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 DyLight®488 antibody (AS09 633, Agrisera; 1:3000). Nuclear DNA was stained with 4’,6-diamidino-2-phenyl-indole (DAPI, 0.4 μg/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; λ ≈ 495 nm) for DyLight-conjugated antibodies and UV-2A filter (UV light; λ ≈ 365 nm) for DAPI. All images were recorded at exactly the same time of integration using DXM 1200 CCD camera.

Courtesy Dr. Dorota Rybaczek, Łódź University, Poland

Additional information

Related products

Background

Background

Programmed cell death (PCD) is a fundamentally important biological process required to maintain the integrity and homeostasis of multicellular organisms during normal development and as a response to adverse environments. Cdc2 kinase, plays a major role in driving the cell cycle. Synonymes (for Arabidopsis thaliana): CDC2A, CDKA-1

Product citations

Selected references Syu et al. (2014). Impacts of size and shape of silver nanoparticles on Arabidopsis plant growth and gene expression. Plant Physiology and Biochemistry 2014. In print.
immunogen:

KLH-conjugated synthetic peptide derived form Zea mays CDC2 sequence P23111, well conserved in other plant species

Reconstitution: For reconstitution add 50 µl of sterile water
Host: Rabbit
Clonality: Polyclonal
Purity: Serum
Format: Lyophilized
Quantity: 50 µl
storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
tested applications: Immunolocalization (IL), Western blot (WB)
recommended dilution: 1 : 500 (IL), 1 : 2000-1 : 10 000 (WB)
calculated | apparent molecular mass [kDa]:

34-36 (Zea mays)

Confirmed reactivity: Arabidopsis thaliana, Hordeum vulgare,Vicia faba, Oryza sativa, Pinus yunnanensi, Zea mays
predicted reactivity: Pisum sativum, Solanum lycopersicum, Triticum aestivum, Pinus contorta, Populus balsamifera, Physcomitrium patens
not reactive in: No confirmed exceptions from predicted reactivity are currently known
Picture (footer): Application example

CDC2 immunolocalization in plant tissue

Seeds of field bean (Vicia faba L. subsp. minor var. Nadwiślański; 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-Gläser). 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 DyLight®488 antibody (AS09 633, Agrisera; 1:3000). Nuclear DNA was stained with 4’,6-diamidino-2-phenyl-indole (DAPI, 0.4 μg/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; λ ≈ 495 nm) for DyLight-conjugated antibodies and UV-2A filter (UV light; λ ≈ 365 nm) for DAPI. All images were recorded at exactly the same time of integration using DXM 1200 CCD camera.

Courtesy Dr. Dorota Rybaczek, Łódź University, Poland

background:

Programmed cell death (PCD) is a fundamentally important biological process required to maintain the integrity and homeostasis of multicellular organisms during normal development and as a response to adverse environments. Cdc2 kinase, plays a major role in driving the cell cycle. Synonymes (for Arabidopsis thaliana): CDC2A, CDKA-1

All references: Syu et al. (2014). Impacts of size and shape of silver nanoparticles on Arabidopsis plant growth and gene expression. Plant Physiology and Biochemistry 2014. In print.

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