DnaK | chloroplast stromal chaperone

PRODUCT INFORMATION IN PDF

product information Background Procaryotic Hsp70 protein family includes DnaK, HscA (Hsc66), HscC (Hsc62). Those proteins are involved in protein folding, cell protection from environmental stress and other functions which are vital for a living cell. Immunogen recombiant DNAK of Chlamydomonas reinhardtii Host Rabbit Clonality Polyclonal Clone 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 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) Related products AS07 271 | anti-DnaJ | prokaryotic heat shock protein collection of antibodies to heat shock proteins collection of antibodies to chloroplastic proteins Additional information

application information
Recommended dilution		1 : 5000 (WB)
Expected | apparent MW		70 kDa
Confirmed reactivity		Chlamydomonas reinhardtii, Synechocystis 6803 motile, Synechocystis 6803 GT (glucose tolerant strain), Synechococcus elongates 7942
Predicted reactivity
Not reactive in		No confirmed exceptions from predicted reactivity are currently known.
Additional information		It is not determined which isoform of DnaK is recognized by this antibody in Arabidopsis thaliana.
Selected references		Göhre et al. (2006). One of Two Alb3 Proteins Is Essential for the Assembly of the Photosystems and for Cell Survival in Chlamydomonas The Plant Cell 18:1454–1466.

Application example



15 µg of Arabidopsis thaliana leaf extract (1), 10 µg of total protein from: Synechocystis 6803 motile (2), Synechocystis 6803 GT (glucose tolerant strain) (3), Synechococcus elongates 7942 (4), Marker - P ierce™ Prestained Protein MW Marker (kat #26612): Total protein was extracted with following buffer: 10 mM Tris HC l, pH 8.0, 0.5% LDS, 4% glycerol, 0.1 mM EDTA were mixed with sample buffer and denatured for 5 min at 95°C. Samples were separated on 10% S DS -PAGE a nd b lo tted 1 h to nitrocellulose membrane (Amersha m Protran) using tank wet transfer (Bio -Rad) in standard transfer buffer in presence of 20% methanol. Transfer of proteins to the membrane was checked using 0,5% Ponceau S staining before the blocking step. Blots were blocked in buffer (2 % lo w -fat milk in 1xPBS, 0,1% Tween) for 1 h at room temperature (RT) with agitation. Blots were incubated in the primary antibody at a dilution of 1 : 5000 for 1 h at RT with agitation. The antibody solutionwas decanted and the blot was rinsed briefly twice, then washed once f or 15 min and 3 times for 5 min in PBS -T at RT with agitation. Blot was incubated in secondary antibody (goat anti-rabbit IgG, AS09 602, Agrisera ) dilut ed to 1 :30 000 in for 1 h at RT with agitation. The blot was was washed as above and developed for 5 min with Clarity Western ECL Substrate and ChemiDoc detection system (Bio-Rad).

Courtesy Dr. Elena Pojidaeva, Laboratory of Plant Gene Expression, Timiryazev Institute of Plant Physiology RAS, 127276 Moscow Russia

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