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HSP23.5 | Heat shock protein 23.5 (mitochondrial)

AS15 2979 | 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) (AS15 2979)

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Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana HSP23.5, UniProt: Q9FGM9, TAIR: AT5G51440

Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum in PBS, pH 7.4
Format Lyophilized
Quantity 50 µg
Reconstitution For reconstitution add 25 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 : 500 (WB)
Expected | apparent MW

23.5 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana recombinant HSP23.5
Predicted reactivity

Arabidopsis thaliana

Not reactive in

Chlamydomonas reinhardtii

Application examples

Application examples application example

western blot using anti-Arabidopsis HSP23.5 antibody

HSP23.5 protein was synthesized in vitro using using the PURExpress In Vitro Protein Synthesis Kit (NEB) according to manufacturer instructions. 20 µL of reaction mixture were separated on 13.5 % SDS-PAGE, and blotted 1h to PVDF (0.2 µm Immobilon PSQ transfer membrane, Millipore) using tank transfer (10 mM CAPS pH 11, 10 % ethanol). Blot was blocked with TBS containing 1.5 % Tween 20 by incubation at room temperature (RT). Blot was briefly rinsed three times with TBS-T (TBS with 0.05 % Tween 20) and incubated overnight at 4°C with the primary antibody at a dilution of 1:1 000 in TBS-T. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 15 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (goat anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:75 000 in TBS-T for 1h at RT with agitation. The blot was washed as above and developed for 2 min with chemiluminescent detection reagent.

Note: Protein was synthesized in vitro using the PURExpress In Vitro Protein Synthesis Kit (NEB)

Courtesty Dr. David Macherel, Mitostress team, IRHS, France.

Additional information

Additional information This antibody is recognizing recombinant HSP23.5 and reactivity on endogenous protein needs to be confirmed. So far HSP23.5 could not be detected in whole seedling extracts under conditions used to detect HSP23.6.

High sensitivity reagents and enrichment are necessary due to low amounts of HSP23.5 in endogenous tissue.

Antibody is not cross reacting with cytosolic HSPs.

Please, note that there might be no HSPs accumulation below temperature of 32-34°C. HSPs are induced when the plant experience temperatures higher than the growing temperature with around 10°C. So, the HSPs induction temperatures for plants grown at 18C differ from these for plants grown at 24C.

Another very effective parameter is the humidity. When using low humidity the plant has a chance to cool down through transpiration. In this case the HSPs induction requires higher temperatures.

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collection of antibodies to plant heat shock proteins

Background

Background

HSP23.5 | heat shock protein 23.5 (mitochondrial) belongs to a small heat shock protein family HSP20.

Product citations

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