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RCD1 | Radical cell death 1

AS12 2602 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

RCD1 | Radical cell death 1 in the group Antibodies for Plant/Algal  / Developmental Biology / Signal transduction at Agrisera AB (Antibodies for research) (AS12 2602)

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Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana RCD1 protein sequence UniProt: Q8RY59, TAIR: AT1G32230

Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum in PBS, pH 7.4
Format Lyophilized in PBS pH 7.4
Quantity 100 ĩg
Reconstitution For reconstitution add 50 ĩl of sterile water.
Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications Western blot (WB) following immunoprecipitation (IP)
Recommended dilution 1 : 1000 (WB) on samples following IP
Expected | apparent MW

65 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana
Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application examples

 application information 

western blot using anti-RCD1 antibodies

5 mg of total protein from Arabidopsis thaliana seedlings expressing RCD1-HA under native promoter extracted with RIPA buffer for protein extraction (50 mM Tris-HCl pH 7,5, 150 mM NaCl, 1% Triton X-100, 0.5% sodium deoxycholate, protease inhibitor cocktail 1:100, 50 uM MG132) were used for immunoprecipitation and afterwards separated on 10% SDS-PAGE and blotted 1h to PVDF using tank transfer. Blots were blocked with 5% milk for 1h at room temperature (RT) with agitation. Blot was incubated in the anti-HA antibody (Roche) at a dilution of 1: 1 000 in 1% milk, TBS-T overnight at +4C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rat IgG horse radish peroxidase conjugated) diluted to 1:10 000 in 1% milk TBS-T for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL Prime, GE Healthcare. Exposure time was 1 min.

Courtesy Dr. Julia Vainonen, Helsinki University, Finland

immunoprecipitation using anti-RCD1 antibodies
5 µl of anti-RCD1 antibody was used for immunoprecipiation. Samples: transgenic line expressing HA-tagged RCD1 (Jaspers et al., 2009, Plant J) (1), rcd1-4 mutant (2). Afterwards these samples were separated on 10% SDS-PAGE and blotted 1h to PVDF using tank transfer. Blots were blocked with 5% milk for 1h at room temperature (RT) with agitation. Blot was incubated in the anti-HA antibody (Roche) at a dilution of 1: 1 000 in 1% milk, TBS-T overnight at +4C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rat IgG horse radish peroxidase conjugated) diluted to 1:10 000 in 1% milk TBS-T for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL Prime, GE Healthcare. Exposure time was 1 min.

Courtesy Dr. Julia Vainonen, Helsinki University, Finland

Additional information

Protein has three splice variants: 63.8 + 65.6 + 65.7 kD

Peptide used to elicit this antibody was designed for detection of all (3) RCD1 splice variants but NOT the close relative SRO1.

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Background

Background RCD1 (Radical cell death 1) functions as a regulator of oxidative-stress hormonal and developmental responses. Alternative name: Inactive poly [ADP-ribose] polymerase RCD1.

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