AB326569

Anti-AGO2(phospho S387) antibody [EPR29226-168]

RabMAb
Recombinant
20ul selling size
What is this?

Be the first to review this product! Submit a review

(0 Publication)

Rabbit Recombinant Monoclonal Argonaute-2 phospho S387 antibody. Suitable for WB, ICC/IF, IP, Dot and reacts with Transfected cell lysate - Human, Transfected cell line - Human, Synthetic peptide - Human samples.

View Alternative Names

EIF2C2, AGO2, Protein argonaute-2, Argonaute2, hAgo2, Argonaute RISC catalytic component 2, Eukaryotic translation initiation factor 2C 2, PAZ Piwi domain protein, Protein slicer, eIF-2C 2, eIF2C 2, PPD

4 Images

Immunocytochemistry/ Immunofluorescence - Anti-AGO2(phospho S387) antibody [EPR29226-168] (AB326569)

ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-AGO2(phospho S387) antibody [EPR29226-168] (AB326569)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells transfected with AKT vector containing a GFP-tag® and AGO2(S387A) vector containing a myc-His-tag® and 293T cells transfected with AKT vector containing a GFP-tag® and AGO2 vector containing a myc-His-tag® labelling AGO2(phospho S387) with ab326569 at 1/50 (10.08 µg/ml) dilution, followed by ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed antibody at 1/1000 dilution (Magenta).

Confocal image showing positive staining in 293T cells transfected with AKT vector containing a GFP-tag® and AGO2 vector containing a myc-His-tag® (shown in magenta), and no staining in 293T cells transfected with AKT vector containing a GFP-tag® and AGO2(S387A) vector containing a myc-His-tag®. GFP was observed in green. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Lab

Immunoprecipitation - Anti-AGO2(phospho S387) antibody [EPR29226-168] (AB326569)

AGO2(phospho S387) was immunoprecipitated from 0.35 mg HEK293T cells were co-transfected with human AGO2 (His-tagged) and human AKT3 (EGFP-tagged) expression vectors, an upstream kinase used to enhance AGO2 phosphorylation at S387, whole cell lysate with ab326569 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab326569 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HEK293T cells were co-transfected with human AGO2 (His-tagged) and human AKT3 (EGFP-tagged) expression vectors, an upstream kinase used to enhance AGO2 phosphorylation at S387, whole cell lysate
Lane 2 : ab326569 IP in HEK293T cells were co-transfected with human AGO2 (His-tagged) and human AKT3 (EGFP-tagged) expression vectors, an upstream kinase used to enhance AGO2 phosphorylation at S387, whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab326569 in HEK293T cells were co-transfected with human AGO2 (His-tagged) and human AKT3 (EGFP-tagged) expression vectors, an upstream kinase used to enhance AGO2 phosphorylation at S387, whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 180 seconds.

All lanes:

Immunoprecipitation - Anti-AGO2(phospho S387) antibody [EPR29226-168] (ab326569) at 1/1000 dilution

Lane 1:

HEK293T cells were co-transfected with human AGO2 (His-tagged) and human AKT3 (EGFP-tagged) expression vectors, an upstream kinase used to enhance AGO2 phosphorylation at S387, whole cell lysate at 10 µg

Lane 2:

ab326569 at 1/30 IP in HEK293T cells were co-transfected with human AGO2 (His-tagged) and human AKT3 (EGFP-tagged) expression vectors, an upstream kinase used to enhance AGO2 phosphorylation at S387, whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab326569 in HEK293T cells were co-transfected with human AGO2 (His-tagged) and human AKT3 (EGFP-tagged) expression vectors, an upstream kinase used to enhance AGO2 phosphorylation at S387, whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 97 kDa

false

Exposure time: 180s

Lab

Western blot - Anti-AGO2(phospho S387) antibody [EPR29226-168] (AB326569)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa); Anti-AGO2 antibody [EPR10411] (ab186733) (1 : 1000) (97KDa); Anti-GFP antibody [ECA-385] (ab32146) (1 : 1000) (75KDa)

All lanes:

Western blot - Anti-AGO2(phospho S387) antibody [EPR29226-168] (ab326569) at 1/1000 dilution

Lane 1:

HEK293T cells were co-transfected with human AGO2(S387A) (His-tagged) and human AKT3 (EGFP-tagged) expression vectors, an upstream kinase used to enhance AGO2 phosphorylation at S387, whole cell lysate (Untreated membrane) at 20 µg

Lane 2:

Lane 3:

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 97 kDa,36 kDa,75 kDa

false

Exposure time: 10s

Lab

Dot Blot - Anti-AGO2(phospho S387) antibody [EPR29226-168] (AB326569)

Dot blot analysis of AGO2(phospho S387) using ab326569 at 1 : 1000 (0.504 µg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane 1 : Human AGO2 non-phospho peptide

Lane 2 : Human AGO2 (phospho S387) peptide a

Lane 3 : Human AGO2 (phospho S387) peptide b

Lane 4 : Human AGO2 (phospho S385) peptide

Lane 5 : Human AGO2 (phospho T390) peptide

Exposure time : 180 seconds.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Dot Blot - Anti-AGO2(phospho S387) antibody [EPR29226-168] (ab326569) at 1/1000 dilution

Lane 1:

Human AGO2 non-phospho peptide

Lane 2:

Human AGO2 (phospho S387) peptide a

Lane 3:

Human AGO2 (phospho S387) peptide b

Lane 4:

Human AGO2 (phospho S385) peptide

Lane 5:

Human AGO2 (phospho T390) peptide

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 180s

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR29226-168

Isotype

IgG

Carrier free

Reacts with

Human

Applications

ICC/IF, Dot, WB, IP

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

style

left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "predicted", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "predicted", "IP-species-dilution-info": "", "IP-species-notes": "", "Dot-species-checked": "predicted", "Dot-species-dilution-info": "", "Dot-species-notes": "" }, "Synthetic peptide - Human": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "1/1000", "Dot-species-notes": "" }, "Transfected cell line - Human": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/50", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "" }, "Transfected cell lysate - Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/30", "IP-species-notes": "", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "" } } }

style

left-column

Product details

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

style

left-column

Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

style

left-column

style

left-column

style

left-column

Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

style

left-column

Target data

Required for RNA-mediated gene silencing (RNAi) by the RNA-induced silencing complex (RISC). The 'minimal RISC' appears to include AGO2 bound to a short guide RNA such as a microRNA (miRNA) or short interfering RNA (siRNA). These guide RNAs direct RISC to complementary mRNAs that are targets for RISC-mediated gene silencing. The precise mechanism of gene silencing depends on the degree of complementarity between the miRNA or siRNA and its target. Binding of RISC to a perfectly complementary mRNA generally results in silencing due to endonucleolytic cleavage of the mRNA specifically by AGO2. Binding of RISC to a partially complementary mRNA results in silencing through inhibition of translation, and this is independent of endonuclease activity. May inhibit translation initiation by binding to the 7-methylguanosine cap, thereby preventing the recruitment of the translation initiation factor eIF4-E. May also inhibit translation initiation via interaction with EIF6, which itself binds to the 60S ribosomal subunit and prevents its association with the 40S ribosomal subunit. The inhibition of translational initiation leads to the accumulation of the affected mRNA in cytoplasmic processing bodies (P-bodies), where mRNA degradation may subsequently occur. In some cases RISC-mediated translational repression is also observed for miRNAs that perfectly match the 3' untranslated region (3'-UTR). Can also up-regulate the translation of specific mRNAs under certain growth conditions. Binds to the AU element of the 3'-UTR of the TNF mRNA and up-regulates translation under conditions of serum starvation. Also required for transcriptional gene silencing (TGS), in which short RNAs known as antigene RNAs or agRNAs direct the transcriptional repression of complementary promoter regions.. (Microbial infection) Upon Sars-CoV-2 infection, associates with viral miRNA-like small RNA, CoV2-miR-O7a, and may repress mRNAs, such as BATF2, to evade the IFN response.

See full target information AGO2 pS387

style

left-column

style

left-column

style

right-column

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com