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HSP23.5 | Heat shock protein 23.5 (mitochondrial)

AS21 4524 | Clonality: Polyclonal |  Host: Rabbit | Reactivity: Arabidopsis thaliana

HSP23.5 |  Heat shock protein 23.5 (mitochondrial)  in the group Antibodies for Plant/Algal  / Environmental Stress / Heat shock at Agrisera AB (Antibodies for research) (AS21 4524)

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Product Information

Immunogen Truncated recombinant HSP23.5 of Arabidopsis thaliana, UniProt: Q9FGM9-1 , TAIR: AT5G51440
Host

Rabbit

Clonality Polyclonal
Purity Antigen affinity purified serum, in PBS pH 7.4
Format Lyophilized
Quantity 50 µg
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 : 1000 (WB)
Expected | apparent MW 23.5 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity Species of your interest not listed? Contact us
Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples Western blot using anti-plant HSP23.5 antibodies

Samples: 

1- 20 μg of Arabidopsis thaliana Col-0 whole protein extract
2- 20 μg of an Arabidopsis thaliana Hsp23.5 null mutant whole protein extract
3- 20 μg of an Arabidopsis thaliana Hsp23.6 null mutant whole protein extract
4- 20 μg of an Arabidopsis thaliana Hsp23.5 and HSp23.6 double null mutant whole protein extract
5- 20 μg of an Arabidopsis thaliana Hsp23.5, 23.6 and 26.5 triple null mutant whole protein extract

Protein for lanes 1-5 was extracted from plants kept at RT

6- 20 μg of Arabidopsis thaliana Col-0 whole protein extract
7- 20 μg of an Arabidopsis thaliana Hsp23.5 null mutant whole protein extract
8- 20 μg of an Arabidopsis thaliana Hsp23.6 null mutant whole protein extract
9 - 20 μg of an Arabidopsis thaliana Hsp23.5 and HSp23.6 double null mutant whole protein extract
10 - 20 μg of an Arabidopsis Hsp23.5, 23.6 and 26.5 triple null mutant whole protein extract

Protein for lanes 6-10 was extracted from plants heat stressed at 38°C for 1 hour, left to recover at RT for 3 hours and then flash frozen.

20 μg/well of total protein extracted from 10 day old Arabidopsis thaliana seedlings (WT and mutant, flash frozen in liquid nitrogen and stored at -85°C) with 60 mM Tris-HCl pH 6.8, 60 mM DTT, 2% SDS, 1.5% Sucrose and 0.05% bromophenol blue and denatured with 1X SDS Loading Dye (10% glycerol, 0.002% bromophenol blue, 0.5% SDS, 50 mM Tris-HCl pH 6.8) at 95°C for 5 min were separated on 4- 20% SDS-PAGE (nUView Tris-Glycine Precast Gel, NuSep) and blotted 1h to nitrocellulose (0.45 μm pore size) using semi-dry transfer. Blot was blocked with 5% milk in TBS-T for 15 minutes, blocking solution was replaced and blocked for another 15 minutes for 30 minutes total with agitation. The blot was incubated in the primary antibody at a dilution of 1:1000 for ~6 hours at 4°C with agitation. The antibody solution was decanted, and the blot was rinsed briefly twice, then washed for 10 minutes, a process repeated three times in total for 30 minutes of washes with TBS-T at RT with agitation. Blot was incubated in Goat Anti- rabbit IgG (horse radish peroxidase conjugated) diluted 1:5000 in TBS-T with agitation. The blot was washed as above and developed for 2 minutes in homemade ECL solution (1.25 mM Luminol, 0.1 M Tris pH 8.5, 0.2 mM p-coumaric acid, 1 mL of solution mixed with 3 μL of 3% hydrogen peroxide just before development). Exposure time was 11 minutes in a G:Box iChemi XT (Syngene).

Courtesy of Fabian Suri-Payer, Vierling Lab,Department of Biochemistry & Molecular Biology University of Massachusetts Amherst, USA

Additional information

Related products

Background

Background HSP23.5 (Heat shock protein 23.5 (mitochondrial) is involved in plant heat shock response. 

Product citations

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