| Application | Recommended Dilution |
|---|---|
| WB | 1:1000-1:5000 |
| IHC | 1:50-1:200 |
| IF | 1:50-1:200 |
Western Blot
Positive WB detected in: A549 whole cell lysate, PC-3 whole cell lysate, HepG2 whole cell lysate, MCF-7 whole cell lysate, 293 whole cell lysate, Hela whole cell lysate
All lanes: PD-L1 antibody at 1:1000
Secondary
Goat polyclonal to Mouse IgG at 1/10000 dilution
Predicted band size: 34, 21 kDa
Observed band size: 55, 70 kDa
IHC image of CSB-MA878942A1m diluted at 1:100 and staining in paraffin-embedded human tonsil tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.
IHC image of CSB-MA878942A1m diluted at 1:100 and staining in paraffin-embedded human colon cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.
Immunofluorescence staining of 293 cells with CSB-MA878942A1m at 1:150, counter-stained with DAPI. The cells were blocked in 10% normal Goat Serum and then incubated with the primary antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L).
Immunofluorescence staining of A549 cells with CSB-MA878942A1m at 1:150, counter-stained with DAPI. The cells were blocked in 10% normal Goat Serum and then incubated with the primary antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L).
Immunofluorescence staining of Hela cells with CSB-MA878942A1m at 1:150, counter-stained with DAPI. The cells were blocked in 10% normal Goat Serum and then incubated with the primary antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L).
Overlay histogram showing 293 cells stained with CSB-MA878942A1m (red line) at 1:300. The cells were incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by primary antibody for 1 h at 4°C. The secondary antibody used was FITC goat anti-mouse IgG(H+L) at 1/200 dilution for 1 h at 4°C. Isotype control antibody (green line) was used under the same conditions. Acquisition of >10,000 events was performed.
Overlay histogram showing A549 cells stained with CSB-MA878942A1m (red line) at 1:300. The cells were incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by primary antibody for 1 h at 4°C. The secondary antibody used was FITC goat anti-mouse IgG(H+L) at 1/200 dilution for 1 h at 4°C. Isotype control antibody (green line) was used under the same conditions. Acquisition of >10,000 events was performed.
Overlay histogram showing Hela cells stained with CSB-MA878942A1m (red line) at 1:300. The cells were incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by primary antibody for 1 h at 4°C. The secondary antibody used was FITC goat anti-mouse IgG(H+L) at 1/200 dilution for 1 h at 4°C. Isotype control antibody (green line) was used under the same conditions. Acquisition of >10,000 events was performed.
In the preparation of this PD-L1 monoclonal antibody, a mouse is selected as the host animal. The recombinant human PD-L1 protein (19-238aa) was injected into the mouse to induce an immune response. Next, the spleen cells were removed from the immunized mouse and then fused with myeloma cells to generate hybridomas. PD-L1 antibody-producing-hybridomas were selected to culture. The PD-L1 monoclonal antibody was extracted from the mouse ascites and underwent purification by protein G affinity chromatography. Its specificity for human PD-L1 protein was validated in multiple applications, including ELISA, WB, IHC, IF, and FC.
PD-L1 plays a key role in regulating the immune system. When PD-L1 binds to its receptor PD-1 on T cells, it inhibits the T cell response and prevents it from attacking PD-L1-expressing cells. This mechanism is often exploited by cancer cells, which can upregulate PD-L1 expression to evade destruction by the immune system. Targeting PD-L1 with immunotherapy drugs has become an important treatment strategy for some types of cancer.
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