Argonaute Antibody Sampler Kit
cat.: HAK21191
Product Type: Antibody Sampler Kit
Species reactivity: Human
Applications: WB
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Uniprot #: SwissProt: Q9UL18 Human | Q8CJG1 Mouse | Q9UKV8 Human | Q8CJG0 Mouse | Q9QZ81 Rat | Q9H9G7 Human | Q8CJF9 Mouse | Q9HCK5 Human
Entrez Gene: 313594 Rat
Unigene: 92025 Rat
Images
HAK21191_1.jpg Fig1: Western blot analysis of AGO1 on different lysates with Rabbit anti-AGO1 antibody (HA724101) at 1/5,000 dilution.

Lane 1: MCF7 cell lysate (20 µg/Lane)
Lane 2: HEK-293 cell lysate (20 µg/Lane)
Lane 3: HeLa cell lysate (20 µg/Lane)
Lane 4: NIH/3T3 cell lysate (20 µg/Lane)
Lane 5: PC-12 cell lysate (20 µg/Lane)
Lane 6: COS-1 cell lysate (20 µg/Lane)
Lane 7: Mouse lung tissue lysate (30 µg/Lane)

Predicted band size: 97 kDa
Observed band size: 97 kDa

Exposure time: 10 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA724101) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21191_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-AGO1 antibody (HA724101) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA724101) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HAK21191_3.jpg Fig3: Immunocytochemistry analysis of C6 cells labeling Argonaute 2 with Rabbit anti-Argonaute 2 antibody (ET1702-39) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Argonaute 2 antibody (ET1702-39) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HAK21191_4.jpg Fig4: Flow cytometric analysis of RAW264.7 cells labeling Argonaute 2.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-39, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HAK21191_5.jpg Fig5: Western blot analysis of AGO4 on different lysates with Rabbit anti-AGO4 antibody (HA500617) at 1/5,000 dilution.

Lane 1: HEK-293 cell lysate
Lane 2: HeLa cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 97 kDa
Observed band size: 97 kDa

Exposure time: 9 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA500617) at 1/5,000 dilution was used in 5% BSA at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.