Western blot (WB): | 1:500-2000 |
Immunohistochemistry (IHC): | 1:50-400 |
Immunofluorescence (IF): | 1:50-400 |
Flow Cytometry (Fixed): | 1:50-200 |
(Boiling the paraffin sections in 10mM citrate buffer,pH6.0,or PH8.0 EDTA repair liquid for 20 mins is required for the staining of formalin/paraffin sections.) Optimal working dilutions must be determined by end user. |
Figure 1. Western blot analysis of anti-CD45/PTPRC antibody (PB9096). The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Jurkat whole cell lysates,
Lane 2: human Raji whole cell lysates.
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-CD45/PTPRC antigen affinity purified polyclonal antibody (PB9096) and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for CD45/PTPRC at approximately 180-250 kDa. The expected band size for CD45/PTPRC is at 147 kDa.
Figure 2. IHC analysis of CD45 using anti-CD45 antibody (PB9096).CD45 was detected in paraffin-embedded section of human tonsil 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 1μg/ml rabbit anti-CD45 Antibody (PB9096) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 3. IF analysis of CD45 using anti- CD45 antibody (PB9096)
CD45 was detected in paraffin-embedded section of human tonsil 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 1μg/mL rabbit anti- CD45 Antibody (PB9096) overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Figure 4. IF analysis of CD45 using anti- CD45 antibody (PB9096)
CD45 was detected in paraffin-embedded section of human tonsil 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 1μg/mL rabbit anti- CD45 Antibody (PB9096) overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Figure 5. Flow Cytometry analysis of THP-1 cells using anti- CD45 antibody (PB9096).
Overlay histogram showing THP-1 cells stained with PB9096 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti- CD45 Antibody (PB9096, 1:100) for 30 min at 20°C. DyLight488 conjugated goat anti-rabbit IgG (BA1127, 1:100) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1:100) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Figure 1. Western blot analysis of anti-CD45/PTPRC antibody (PB9096). The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Jurkat whole cell lysates,
Lane 2: human Raji whole cell lysates.
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-CD45/PTPRC antigen affinity purified polyclonal antibody (PB9096) and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for CD45/PTPRC at approximately 180-250 kDa. The expected band size for CD45/PTPRC is at 147 kDa.
Figure 2. IHC analysis of CD45 using anti-CD45 antibody (PB9096).CD45 was detected in paraffin-embedded section of human tonsil 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 1μg/ml rabbit anti-CD45 Antibody (PB9096) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 3. IF analysis of CD45 using anti- CD45 antibody (PB9096)
CD45 was detected in paraffin-embedded section of human tonsil 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 1μg/mL rabbit anti- CD45 Antibody (PB9096) overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Figure 4. IF analysis of CD45 using anti- CD45 antibody (PB9096)
CD45 was detected in paraffin-embedded section of human tonsil 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 1μg/mL rabbit anti- CD45 Antibody (PB9096) overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Figure 5. Flow Cytometry analysis of THP-1 cells using anti- CD45 antibody (PB9096).
Overlay histogram showing THP-1 cells stained with PB9096 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti- CD45 Antibody (PB9096, 1:100) for 30 min at 20°C. DyLight488 conjugated goat anti-rabbit IgG (BA1127, 1:100) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1:100) used under the same conditions. Unlabelled sample (Red line) was also used as a control.