PNP Antibody – Rabbit Polyclonal

Figure 1. Western blot analysis of PNP using anti-PNP antibody (MBS178628).
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: rat thymus tissue lysates,
Lane 2: rat ovary tissue lysates,
Lane 3: mouse liver tissue lysates,
Lane 4: human placneta 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-PNP antigen affinity purified polyclonal antibody at 0.5ug/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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for PNP at approximately 38KD. The expected band size for PNP is at 38KD.

Figure 2. IHC analysis of PNP using anti-PNP antibody (MBS178628).
PNP was detected in paraffin-embedded section of mouse kidney tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-PNP Antibody (MBS178628) 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) with DAB as the chromogen.

Figure 3. IHC analysis of PNP using anti-PNP antibody (MBS178628).
PNP was detected in paraffin-embedded section of rat kidney tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-PNP Antibody (MBS178628) 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) with DAB as the chromogen.

Figure 4. IHC analysis of PNP using anti-PNP antibody (MBS178628).
PNP was detected in paraffin-embedded section of human liver cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-PNP Antibody (MBS178628) 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) with DAB as the chromogen.

Figure 5. IHC analysis of PNP using anti-PNP antibody (MBS178628).
PNP was detected in paraffin-embedded section of human renal cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-PNP Antibody (MBS178628) 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) with DAB as the chromogen.

Figure 6. Flow Cytometry analysis of U937 cells using anti-PNP antibody (MBS178628).
Overlay histogram showing U937 cells stained with MBS178628 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PNP Antibody (MBS178628,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight ®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 7. Flow Cytometry analysis of U251 cells using anti-PNP antibody (MBS178628).
Overlay histogram showing U251 cells stained with MBS178628 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PNP Antibody (MBS178628,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight ®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.