PYK10 | Beta-Galactosidase (internal) (ER marker, immunolocalisation)
AS20 4409 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana | Marker antibody for ER in immunolocalisation
DATA SHEET IN PDF| Data sheet | Product citations | Protocols | Add review |
Product Information
Immunogen
Conjugated peptide, derived from Arabidopsis thaliana internal part of PYK10, UniProt: A0A178VCN3, TAIR: At3g08880.
Host
Rabbit
Clonality
Polyclonal
Purity
Total IgG, purified on Protein A
Format
Liquid at 2 mg/ml in PBS, 50% glycerol. Filter sterilized. No preservative or carrier protein.
Quantity
100 µg
Storage
Store at -20°C; once 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 material adhering to the cap or sides of the tubes.
Tested applications
Immunofluorescence (IF), Immunohistochemistry (IHC), Immunoprecipitation (IP), Western blot (WB)
Recommended dilution
1: 100 -1: 500 (IF), 1: 500 (IHC), 1: 500 (IP), 1: 5000 - 1: 20 000 (WB)
Expected | apparent MW
59.7 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
Arabidopsis thaliana 7 day-old seedlings was freshly extracted with 2x SDS-sample buffer (+ 2ME) for SDS-PAGE and denatured with 4X SDS buffer at 95°C for 5 min. were separated on 12.5 % SDS-PAGE and blotted to PVDF membrane in semi-dry system. Blot was blocked with 5 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5000 in TBS-T for 1h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendations.
Left panel: anti-PYK10 antibodies, middle panel: GFP, right panel: marged
The P1 pellet was obtained from 8 day-old seedlings (carrying GFP gene with ER-retension signal). Anti-PYK10 antibodies: 1: 500, secondary antibodies: goat anti-rabbit IgG Alexa Fluor546 at 1: 1000.
PYK10 is localising to ER bodies,
Arabidopsis thaliana 7 day-old seedlings was freshly extracted with 2x SDS-sample buffer (+ 2ME) for SDS-PAGE and denatured with 4X SDS buffer at 95°C for 5 min. were separated on 12.5 % SDS-PAGE and blotted to PVDF membrane in semi-dry system. Blot was blocked with 5 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5000 in TBS-T for 1h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendations.
Left panel: anti-PYK10 antibodies, middle panel: GFP, right panel: marged
The P1 pellet was obtained from 8 day-old seedlings (carrying GFP gene with ER-retension signal). Anti-PYK10 antibodies: 1: 500, secondary antibodies: goat anti-rabbit IgG Alexa Fluor546 at 1: 1000.
PYK10 is localising to ER bodies,
Additional information
N-terminal signal peptide including 24 amino acis and ER retention signal is removed from the mature protein.
Background
Background
PYK10 is the main component of ER bodies. It has hydrolase activity, hydrolyzing O-glycosyl compounds It may produce defense compounds when plants are damaged by insects or wounding.
Product citations
Selected references
Yamada et al. (2013). Identification of Two Novel Endoplasmic Reticulum Body-Specific Integral Membrane Proteins. Plant Physiol. 2013 Jan;161(1):108-20. doi: 10.1104/pp.112.207654. (Western blot, Arabidopsis thaliana)
Nagano et al. (2005). Activation of an ER-body-localized ß-Glucosidase via a Cytosolic Binding Partner in Damaged Tissues of Arabidopsis thaliana. Plant Cell Physiol. 2005 Jul;46(7):1140-8. doi: 10.1093/pcp/pci126. (Immunoprecipiation, Western blot, Arabidopsis thaliana)
Matshushima et al. (2003). A novel ER-derived compartment, the ER body, selectively accumulates a beta-glucosidase with an ER-retention signal in Arabidopsis. Plant J . 2003 Feb;33(3):493-502. doi: 10.1046/j.1365-313x.2003.01636.x. Immunofluorescence, Immunohistochemistry, Western blot, Arabidopsis thaliana)
Nagano et al. (2005). Activation of an ER-body-localized ß-Glucosidase via a Cytosolic Binding Partner in Damaged Tissues of Arabidopsis thaliana. Plant Cell Physiol. 2005 Jul;46(7):1140-8. doi: 10.1093/pcp/pci126. (Immunoprecipiation, Western blot, Arabidopsis thaliana)
Matshushima et al. (2003). A novel ER-derived compartment, the ER body, selectively accumulates a beta-glucosidase with an ER-retention signal in Arabidopsis. Plant J . 2003 Feb;33(3):493-502. doi: 10.1046/j.1365-313x.2003.01636.x. Immunofluorescence, Immunohistochemistry, Western blot, Arabidopsis thaliana)
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