AGO10 | Argonaute 10

Name: AGO10 | Argonaute 10
SKU: AS15 3071
Price: 302 EUR
Availability: InStock

AS15 3071 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

AGO10 | Argonaute 10 in the group Antibodies Plant/Algal / DNA/RNA/Cell Cycle / plant RNA at Agrisera AB (Antibodies for research) (AS15 3071)

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana AGO10 protein sequence, Uniprot: Q9XGW1, TAIR: AT5G43810

Host Rabbit

Clonality Polyclonal

Purity Immunogen 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 the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.

Tested applications Western blot (WB)

Recommended dilution 1 : 10 000 (WB)

Expected | apparent MW 110,9 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana

Predicted reactivity

A. lyrata, B. napus, C. rubella, C. clementina, C. sinensis, E. salsugineum, G. arboreum, G. raimondii. N. benthamiana

Species of your interest not listed? Contact us

Not reactive in

Zea mays

Application examples

Application examples Application example

50 µg of total protein from Arabidopsis thaliana inflorescences were extracted with extraction buffer (50 mM Tris pH7.5; 150 mM NaCl; 1 mM EDTA; 10 % v/v Glycerin; 1 mM DTT, 1x Complete Protease Inhibitor Cocktail, Roche) and denatured with Laemmli buffer at 95ºC 5 min. were separated on 10% SDS-PAGE and blotted 1.5 h to PVDF using tank transfer. Blots were blocked with blocking buffer (3% milk powder; 1x TBS; 0.1% Tween-20) 1 h at RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:10 000 ON at 4ºC with agitation. The antibody solution was decanted and the blot was rinsed briefly and then washed tree times for 15 min. in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:20 000 in blocking buffer for 1h at RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescent detection reagent of extreme femtogram sensitivity, exposed to Amersham Hyperfilms ECL for 5 minutes. ago10-2 mutant is described here.

Courtesy of Dr. Dr. Pablo Manavella, Instituto de Agrobiotecnología del Litoral (IAL), Argentina

Additional information

AGO expression may be cell/tissue specific and using floral tissue is recommended where most of the AGOs are expressed the highest. Seedlings can be used as a negative control.

Use of proteasome inhibitors as MG132 can help to stabilize AGO proteins during extraction procedure.

Background

AGO10 is involved in miRNA binding, and in RNA-mediated posttranscriptional gene silencing (PTGS).

Product citations

Selected references Sun et al. (2021) The epigenetic factor FVE orchestrates cytoplasmic SGS3-DRB4-DCL4 activities to promote transgene silencing in Arabidopsis. Sci Adv. 2021 Aug 4;7(32):eabf3898. doi: 10.1126/sciadv.abf3898. PMID: 34348894; PMCID: PMC8336953.
Oliver & Martinez. (2021) Accumulation dynamics of ARGONAUTE proteins during meiosis in Arabidopsis. Plant Reprod. 2021 Nov 23. doi: 10.1007/s00497-021-00434-z. Epub ahead of print. PMID: 34812935.
Sprunck et al. (2019). Elucidating small RNA pathways in Arabidopsis thaliana egg cells.

immunogen:	KLH-conjugated synthetic peptide derived from Arabidopsis thaliana AGO10 protein sequence, Uniprot: Q9XGW1, TAIR: AT5G43810
Reconstitution:	For reconstitution add 50 µl of sterile water
Host:	Rabbit
Clonality:	Polyclonal
Purity:	Immunogen affinity purified serum in PBS pH 7.4.
Format:	Lyophilized
Quantity:	50 µg
storage:	Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
tested applications:	Western blot (WB)
recommended dilution:	1 : 10 000 (WB)
calculated \| apparent molecular mass [kDa]:	110,9 kDa
Confirmed reactivity:	Arabidopsis thaliana
predicted reactivity:	A. lyrata, B. napus, C. rubella, C. clementina, C. sinensis, E. salsugineum, G. arboreum, G. raimondii. N. benthamiana Species of your interest not listed? Contact us
not reactive in:	Zea mays
Picture (footer):	Application example 50 µg of total protein from Arabidopsis thaliana inflorescences were extracted with extraction buffer (50 mM Tris pH7.5; 150 mM NaCl; 1 mM EDTA; 10 % v/v Glycerin; 1 mM DTT, 1x Complete Protease Inhibitor Cocktail, Roche) and denatured with Laemmli buffer at 95ºC 5 min. were separated on 10% SDS-PAGE and blotted 1.5 h to PVDF using tank transfer. Blots were blocked with blocking buffer (3% milk powder; 1x TBS; 0.1% Tween-20) 1 h at RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:10 000 ON at 4ºC with agitation. The antibody solution was decanted and the blot was rinsed briefly and then washed tree times for 15 min. in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:20 000 in blocking buffer for 1h at RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescent detection reagent of extreme femtogram sensitivity, exposed to Amersham Hyperfilms ECL for 5 minutes. ago10-2 mutant is described here. Courtesy of Dr. Dr. Pablo Manavella, Instituto de Agrobiotecnología del Litoral (IAL), Argentina
additional information (application):	AGO expression may be cell/tissue specific and using floral tissue is recommended where most of the AGOs are expressed the highest. Seedlings can be used as a negative control. Use of proteasome inhibitors as MG132 can help to stabilize AGO proteins during extraction procedure.
background:	AGO10 is involved in miRNA binding, and in RNA-mediated posttranscriptional gene silencing (PTGS).
All references:	Sun et al. (2021) The epigenetic factor FVE orchestrates cytoplasmic SGS3-DRB4-DCL4 activities to promote transgene silencing in Arabidopsis. Sci Adv. 2021 Aug 4;7(32):eabf3898. doi: 10.1126/sciadv.abf3898. PMID: 34348894; PMCID: PMC8336953. Oliver & Martinez. (2021) Accumulation dynamics of ARGONAUTE proteins during meiosis in Arabidopsis. Plant Reprod. 2021 Nov 23. doi: 10.1007/s00497-021-00434-z. Epub ahead of print. PMID: 34812935. Sprunck et al. (2019). Elucidating small RNA pathways in Arabidopsis thaliana egg cells.