| Application | Recommended Dilution |
|---|---|
| WB:1 | 5000-160000 |
| IHC | 1:50-1:200 |
| IF | 1:50-1:200 |
| FC | 1:50-1:200 |
| IP | 1µl-4µl |
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
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
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
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.
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.
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.
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).
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).
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).
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.
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.
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.
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
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
The YWHAZ monoclonal antibody recognizes the YWHAZ protein and has been shown to be highly specific in human, mouse, and rat samples. It is suitable for use in a range of applications, including ELISA, WB, IHC, IF, and FC. This YWHAZ monoclonal antibody was obtained from the YWHAZ antibody-secreting hybridomas, which were formed by the fusion of myeloma cells with spleen cells from an immunized mouse. The recombinant human 14-3-3 protein zeta/delta protein (133-212aa) was used to immunize the mouse. This YWHAZ monoclonal antibody undergoes protein A-mediated purification, reaching up to 95% in purity.
YWHAZ, also known as 14-3-3 protein zeta/delta, mainly regulates a wide range of cellular processes such as signal transduction, cell cycle regulation, apoptosis, and gene expression through interactions with various protein partners. YWHAZ is involved in regulating multiple signaling pathways including the PI3K/Akt, MAPK, and NF-κB pathways, and plays an important role in mediating cell survival and growth.
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