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Western Blot
Positive WB detected in: NIH/3T3 whole cell lysate, Hela whole cell lysate, HepG2 whole cell lysate, A549 whole cell lysate, RAW264.7 whole cell lysate, Rat Brain tissue
All lanes YWHAZ antibody at 1:5000
Secondary
Goat polyclonal to mouse IgG at 1/50000 dilution
Predicted band size: 34 KDa
Observed band size: 34 KDa
Exposure time:1min
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Western Blot
Positive WB detected in: Hela whole cell lysate at 20μg, 10μg, 5μg, 2.5μg, 1.25μg, 0.625μg
All lanes: YWHAZ antibody at 1:5000
Secondary
Goat polyclonal to mouse IgG at 1/50000 dilution
Predicted band size: 34 KDa
Observed band size: 34 KDa
Exposure time:5min
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Western Blot
Positive WB detected in: 20μg Hela whole cell lysate YWHAZ antibody at1:5000,1:10000,1:20000,1:40000,1:80000,1:160000
Secondary
Goat polyclonal to mouse IgG at 1/50000 dilution
Predicted band size: 34 KDa
Observed band size: 34 KDa
Exposure time:5min
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IHC image of CSB-MA026293A0m diluted at 1:200 and staining in paraffin-embedded human lung cancer 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 37°C. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit IgG labeled by HRP and visualized using 0.05% DAB.
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IHC image of CSB-MA026293A0m diluted at 1:200 and staining in paraffin-embedded human liver cancer 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 37°C. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit IgG labeled by HRP and visualized using 0.05% DAB.
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IHC image of CSB-MA026293A0m diluted at 1:200 and staining in paraffin-embedded human breast cancer 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 37°C. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit IgG labeled by HRP and visualized using 0.05% DAB.
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Immunofluorescence staining of A549 cells with CSB-MA026293A0m at 1:50, counter-stained with DAPI. The cells were fixed in 4% formaldehyde and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. Nuclear DNA was labeled in blue with DAPI. The secondary antibody was FITC-conjugated AffiniPure Goat Anti-Mouse IgG (H+L).
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Immunofluorescence staining of Hela cells with CSB-MA026293A0m at 1:50, counter-stained with DAPI. The cells were fixed in 4% formaldehyde and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. Nuclear DNA was labeled in blue with DAPI. The secondary antibody was FITC-conjugated AffiniPure Goat Anti-Mouse IgG (H+L).
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Immunofluorescence staining of U251 cells with CSB-MA026293A0m at 1:50, counter-stained with DAPI. The cells were fixed in 4% formaldehyde and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. Nuclear DNA was labeled in blue with DAPI. The secondary antibody was FITC-conjugated AffiniPure Goat Anti-Mouse IgG (H+L).
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Overlay histogram showing A549 cells stained with CSB-MA026293A0m (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100. Then 10% normal goat serum was Incubated to block non-specific protein-protein interactions followed by the antibody (1µg/1*106cells) for 1 h at 4°C. The secondary antibody used was FITC-conjugated Goat Anti-Mouse IgG(H+L) at 1/100 dilution for 30min at 4°C. Isotype control antibody (green line) was mouse IgG2b (1µg/1*106cells) used under the same conditions. Acquisition of >10,000 events was performed.
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Overlay histogram showing Hela cells stained with CSB-MA026293A0m (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100. Then 10% normal goat serum was Incubated to block non-specific protein-protein interactions followed by the antibody (1µg/1*106cells) for 1 h at 4°C. The secondary antibody used was FITC-conjugated Goat Anti-Mouse IgG(H+L) at 1/100 dilution for 30min at 4°C. Isotype control antibody (green line) was mouse IgG2b (1µg/1*106cells) used under the same conditions. Acquisition of >10,000 events was performed.
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Overlay histogram showing HepG2 cells stained with CSB-MA026293A0m (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100. Then 10% normal goat serum was Incubated to block non-specific protein-protein interactions followed by the antibody (1µg/1*106cells) for 1 h at 4°C. The secondary antibody used was FITC-conjugated Goat Anti-Mouse IgG(H+L) at 1/100 dilution for 30min at 4°C. Isotype control antibody (green line) was mouse IgG2b (1µg/1*106cells) used under the same conditions. Acquisition of >10,000 events was performed.
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Immunoprecipitating YWHAZ in A549 whole cell lysate
Lane 1: Mouse control IgG2b instead of CSB-MA026293A0m in A549 whole cell lysate.
Lane 2: CSB-MA026293A0m (1µg) + A549 whole cell lysate (500µg)
Lane 3: A549 whole cell lysate (20µg)
For western blotting, the blot was detected with CSB-MA026293A0m at 1:2000, and a HRP-conjugated Protein G antibody was used as the secondary antibody at 1:50000
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Immunoprecipitating YWHAZ in HepG2 whole cell lysate
Lane 1: Mouse control IgG2b instead of CSB-MA026293A0m in HepG2 whole cell lysate.
Lane 2: CSB-MA026293A0m (1µg) + HepG2 whole cell lysate (500µg)
Lane 3: HepG2 whole cell lysate (20µg)
For western blotting, the blot was detected with CSB-MA026293A0m at 1:2000, and a HRP-conjugated Protein G antibody was used as the secondary antibody at 1:50000
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