AGO9 | Argonaute 9

Name: AGO9 | Argonaute 9
SKU: AS10 673
Price: 424 EUR
Availability: InStock

AS10 673 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

LIMITED STOCK

AGO9 | Argonaute 9 in the group Antibodies Plant/Algal / DNA/RNA/Cell Cycle / plant RNA at Agrisera AB (Antibodies for research) (AS10 673)

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana AGO9 protein sequence UniProt: Q84YI4, TAIR:At5g21150.

Host Rabbit

Clonality Polyclonal

Purity Immunogen affinity purified serum in PBS pH 7.4.

Format Lyophilized

Quantity 100 µg

Reconstitution For reconstitution add 100 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 Immunolocalization, whole-mount (IL), Immunoprecipitation (IP), Western blot (WB)

Recommended dilution 5 µg of antibody per 1 gram of a fresh tissue (IP), 1 : 10 000 (WB)

Expected | apparent MW

101 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana

Predicted reactivity

Arabidopsis thaliana

Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application example

western blot using Arabidopsis AGO9 antibodies

80 µg of Arabidopsis thaliana soluble total cell extract (extracted in 20 mMTris pH7.5, 5mM MgCl2, 2.5mM DTT, 300mM NaCl, 0.1% NP-40, 1% proteaseinhibitor) was separated on 6% SDS-PAGE and blotted 1h to nitrocellulose. Filters were blocked 1h with 5% low-fat milk powder in TBS-TT (0.25% TWEEN20; 0.1% Triton-X) and probed with anti-AGO9 antibody (1:10 000, 1h) and secondary anti-rabbit antibody (HRP conjugated, Agrisera AS09 602) (1:15 000, 1 h) in TBS-TT containing 5% low fat milk powder. Antibody incubations were followed by washings in TBS-TT. All steps were performed at RT with agitation. Blots were developed for 5 min with chemiluminescent detection reagent according the manufacturer's instructions. Exposure time was 30 seconds.

Courtesy Dr. Ericka Havecker, University of Cambridge, UK

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.

A recommended whole-mount immunolocalization protocol can be found here.

Background

AGO9 belongs to a group of argonaute proteins which are catalytic component of the RNA-incudes silencing complex (RISC).

Product citations

Selected references Hou et al. (2021) High-throughput single-cell transcriptomics reveals the female germline differentiation trajectory in Arabidopsis thaliana. Commun Biol. 2021 Oct 1;4(1):1149. doi: 10.1038/s42003-021-02676-z. PMID: 34599277; PMCID: PMC8486858. (immunolocalization)
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. http://dx.doi.org/10.1101/525956
Su et al. (2017). The THO Complex Non-Cell-Autonomously Represses Female Germline Specification through the TAS3-ARF3 Module. Curr Biol. 2017 Jun 5;27(11):1597-1609.e2. doi: 10.1016/j.cub.2017.05.021.
Havecker et al. (2010) The RNA-directed DNA methylation Arabidopsis Argonautes functionally diverge based on expression and interaction with target loci. Plant Cell 22(2): 321-34.

immunogen:	KLH-conjugated synthetic peptide derived from Arabidopsis thaliana AGO9 protein sequence UniProt: Q84YI4, TAIR:At5g21150.
Reconstitution:	For reconstitution add 100 µl of sterile water
Host:	Rabbit
Clonality:	Polyclonal
Purity:	Immunogen affinity purified serum in PBS pH 7.4.
Format:	Lyophilized
Quantity:	100 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:	Immunolocalization, whole-mount (IL), Immunoprecipitation (IP), Western blot (WB)
recommended dilution:	5 µg of antibody per 1 gram of a fresh tissue (IP), 1 : 10 000 (WB)
calculated \| apparent molecular mass [kDa]:	101 kDa
Confirmed reactivity:	Arabidopsis thaliana
predicted reactivity:	Arabidopsis thaliana
not reactive in:	No confirmed exceptions from predicted reactivity are currently known
Picture (footer):	Application example 80 µg of Arabidopsis thaliana soluble total cell extract (extracted in 20 mMTris pH7.5, 5mM MgCl2, 2.5mM DTT, 300mM NaCl, 0.1% NP-40, 1% proteaseinhibitor) was separated on 6% SDS-PAGE and blotted 1h to nitrocellulose. Filters were blocked 1h with 5% low-fat milk powder in TBS-TT (0.25% TWEEN20; 0.1% Triton-X) and probed with anti-AGO9 antibody (1:10 000, 1h) and secondary anti-rabbit antibody (HRP conjugated, Agrisera AS09 602) (1:15 000, 1 h) in TBS-TT containing 5% low fat milk powder. Antibody incubations were followed by washings in TBS-TT. All steps were performed at RT with agitation. Blots were developed for 5 min with chemiluminescent detection reagent according the manufacturer's instructions. Exposure time was 30 seconds. Courtesy Dr. Ericka Havecker, University of Cambridge, UK
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. A recommended whole-mount immunolocalization protocol can be found here.
background:	AGO9 belongs to a group of argonaute proteins which are catalytic component of the RNA-incudes silencing complex (RISC).
All references:	Hou et al. (2021) High-throughput single-cell transcriptomics reveals the female germline differentiation trajectory in Arabidopsis thaliana. Commun Biol. 2021 Oct 1;4(1):1149. doi: 10.1038/s42003-021-02676-z. PMID: 34599277; PMCID: PMC8486858. (immunolocalization) 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. http://dx.doi.org/10.1101/525956 Su et al. (2017). The THO Complex Non-Cell-Autonomously Represses Female Germline Specification through the TAS3-ARF3 Module. Curr Biol. 2017 Jun 5;27(11):1597-1609.e2. doi: 10.1016/j.cub.2017.05.021. Havecker et al. (2010) The RNA-directed DNA methylation Arabidopsis Argonautes functionally diverge based on expression and interaction with target loci. Plant Cell 22(2): 321-34.