YTHDF2 Antibody – Rabbit Polyclonal

Figure 1. Western blot analysis of YTHDF2 using anti-YTHDF2 antibody (MBS1753540).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human K562 whole cell lysates
Lane 2: human Raji whole cell lysates
Lane 3: human HL-60 whole cell lysates
Lane 4: human Hela whole cell lysates
Lane 5: human A549 whole cell lysates
Lane 6: human Hepg2 whole cell lysates
Lane 7: human THP-1 whole cell lysates
Lane 8: rat liver tissue lysates
Lane 9: mouse testis tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-YTHDF2 antigen affinity purified polyclonal antibody (Catalog # MBS1753540) at 0. 25 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for YTHDF2 at approximately 65KD. The expected band size for YTHDF2 is at 65KD.

Figure 2. IHC analysis of YTHDF2 using anti-YTHDF2 antibody (MBS1753540).
YTHDF2 was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-YTHDF2 Antibody (MBS1753540) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen.

Figure 3. IHC analysis of YTHDF2 using anti-YTHDF2 antibody (MBS1753540).
YTHDF2 was detected in paraffin-embedded section of mouse intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-YTHDF2 Antibody (MBS1753540) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen.

Figure 4. IHC analysis of YTHDF2 using anti-YTHDF2 antibody (MBS1753540).
YTHDF2 was detected in paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-YTHDF2 Antibody (MBS1753540) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen.

Figure 5. IF analysis of YTHDF2 using anti- YTHDF2 antibody (MBS1753540).
YTHDF2 was detected in immunocytochemical section of Hep cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti- YTHDF2 Antibody (MBS1753540) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 6. Flow Cytometry analysis of RAW264. 7 cells using anti-YTHDF2 antibody (MBS1753540).
Overlay histogram showing RAW264. 7 cells stained with MBS1753540 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-YTHDF2 Antibody (MBS1753540, 1μg/1x10⁶ cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 7. Flow Cytometry analysis of RH-35 cells using anti-YTHDF2 antibody (MBS1753540).
Overlay histogram showing RH-35 cells stained with MBS1753540 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-YTHDF2 Antibody (MBS1753540, 1μg/1x10⁶ cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 8. Flow Cytometry analysis of THP-1 cells using anti-YTHDF2 antibody (MBS1753540).
Overlay histogram showing THP-1 cells stained with MBS1753540 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-YTHDF2 Antibody (MBS1753540, 1μg/1x10⁶ cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.