AGO1 | Argonaute 1 (Chlamydomonas)
AS14 2776 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Chlamydomonas reinhardii
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Product Information
Immunogen
recombinant AGO1 of Chlamydomonas reinhardtii
Host
Rabbit
Clonality
Polyclonal
Purity
Total IgG. Protein G purified in PBS pH 7.4.
Format
Lyophilized
Quantity
1 mg
Reconstitution
For reconstitution add 50 ĩl 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 : 1000 (WB)
Expected | apparent MW
116,4 | 130 kDa
Reactivity
Confirmed reactivity
Recombinant AGO1 of Chlamydomonas reinhardtii
Application examples
Application examples
Application example
Molecular weight markers (1), recombinant AGO1 of Chlamydomonas reinhardtii (2), recombinant AGO1 of Chlamydomonas reinhardtii (1/2 dilution) (3). Recombinant protein was purified using Ni-NTA Agarose (Qiagen), eluted in 50mM NaH2PO4, 500 mM NaCl and 250 mM imidazole (pH 7.5) and diluted in 1xPBS with 2% SDS. Samples were denatured prior to loading with 5 x loading buffer (350 mM Tris-Cl (pH 7.5), 30% glycerol, 10% SDS and 0.6 M DTT) by heating at 95°C for 5 mins. Samples were separated on NuPAGE 7% Tris-Acetate Protein Gel and blotted 1h to PVDF using tank transfer. Blots were blocked with 1xTBS with 0.1% Tween-20 (TBS-T) and 5% milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 for 15 hrs at 4°C with agitation in TBS-T. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (goat anti-rabbit Ab) diluted to 1:5000 in TBS-T for 1h at RT with agitation. The blot was washed as above and imaged using the Odyssey Infrared Imager and Image Studio v 2.1.
Courtesy of PhD candidate Claire Agius, University of Cambridge, UK
Molecular weight markers (1), recombinant AGO1 of Chlamydomonas reinhardtii (2), recombinant AGO1 of Chlamydomonas reinhardtii (1/2 dilution) (3). Recombinant protein was purified using Ni-NTA Agarose (Qiagen), eluted in 50mM NaH2PO4, 500 mM NaCl and 250 mM imidazole (pH 7.5) and diluted in 1xPBS with 2% SDS. Samples were denatured prior to loading with 5 x loading buffer (350 mM Tris-Cl (pH 7.5), 30% glycerol, 10% SDS and 0.6 M DTT) by heating at 95°C for 5 mins. Samples were separated on NuPAGE 7% Tris-Acetate Protein Gel and blotted 1h to PVDF using tank transfer. Blots were blocked with 1xTBS with 0.1% Tween-20 (TBS-T) and 5% milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 for 15 hrs at 4°C with agitation in TBS-T. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (goat anti-rabbit Ab) diluted to 1:5000 in TBS-T for 1h at RT with agitation. The blot was washed as above and imaged using the Odyssey Infrared Imager and Image Studio v 2.1.
Courtesy of PhD candidate Claire Agius, University of Cambridge, UK
Additional information
Antibody binds to recombinat AGO1 of Chlamydomonas reinhardtii. Endogenous detection remains to be confirmed.
Background
Background
AGO1 belongs to a group of argonaute proteins which are catalytic component of the RNA-incudes silencing complex (RISC). This protein complex is responsible for the gene silencing (RNAi).
Product citations
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