DnaK | chloroplast stromal chaperone
AS07 270 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Chlamydomonas reinhardtii, Synechocystis 6803 motile, Synechocystis 6803 GT (glucose tolerant strain), Synechococcus elongates 7942
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Product Information
Immunogen
recombiant DNAK of Chlamydomonas reinhardtii
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 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)
Recommended dilution
1 : 5000 (WB)
Expected | apparent MW
70 kDa
Reactivity
Confirmed reactivity
Chlamydomonas reinhardtii, Synechocystis 6803 motile, Synechocystis 6803 GT (glucose tolerant strain), Synechococcus elongates sp. PCC7942
Not reactive in
No confirmed exceptions from predicted reactivity are currently known.
Application examples
Application examples
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
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
Additional information
It is not determined which isoform of DnaK is recognized by this antibody in Arabidopsis thaliana.
Background
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.
Product citations
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.
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