Total protein was isolated using RIPA lysis buffer (JRDUN, Shanghai, China). 25 μg protein of each sample was fractionated and transferred onto PVDF nitrocellulose membrane (HATF00010, Millipore, USA) for 12 h. The transfer method was semidry at 25 V on 100 mA current, lasted for 30 minutes, and the pore size of the PVDF membrane was 0.22 μm. Then, the membranes were probed with the primary antibodies overnight at 4°C, followed by the appropriate HRP-conjugated goat anti-rabbit IgG (A0208, Beyotime, China). Protein signals were analyzed using a chemiluminescence system. The antibodies details were provided as follows: TRIM37 (Ab95997, Abcam, UK, 1 : 1000), TRAF2 (Ab60169, Abcam, UK, 1 : 1500), SR-B1(Ab52629, Abcam, UK, 1 : 1000), ABCA1 (Ab18180, Abcam, UK, 1 : 1000), ABCG1 (Ab52617, Abcam, UK, 1 : 2000), NF-KB (#8242, CST, USA, 1 : 1000), H3 (#4499, CST, USA, 1 : 1000), and β-actin (#4970, CST, USA, 1 : 1000).
Ab95997
Manufactured by Abcam
Sourced in United Kingdom
Anti-PHLPP1 antibody (ab95997) is a rabbit polyclonal antibody that recognizes the PHLPP1 (PH domain and leucine rich repeat protein phosphatase 1) protein. PHLPP1 is a serine/threonine protein phosphatase that negatively regulates the AKT signaling pathway. The antibody can be used in various applications such as Western blotting, immunoprecipitation, and immunohistochemistry to detect the PHLPP1 protein.
Automatically generated - may contain errors
Lab products found in correlation
Formalin, by Thermo Fisher Scientific (2 mentions)
Ab17721, by Abcam (2 mentions)
Ab90996, by Abcam (2 mentions)
A11037, by Abcam (2 mentions)
Ab199428, by Abcam (2 mentions)
Hatf00010, by Merck Group (1 mentions)
Ab60169, by Abcam (1 mentions)
Ab52617, by Abcam (1 mentions)
Ab52629, by Abcam (1 mentions)
Ab18180, by Abcam (1 mentions)
Hrp conjugated goat anti rabbit igg, by Beyotime (1 mentions)
3 protocols using ab95997
1
Western Blot Protein Analysis Protocol
2
Immunofluorescence Staining of Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MCF7 and T47D cells were seeded on slides at an appropriate density for 24h. After paraformaldehyde fixation, cells on slides were treated with 0.3% Triton X-100 for permeabilization of the plasma membrane and then immunostained with specific antibodies including anti-LSD1 (Abcam, ab90996 1:200 or ab17721; 1:200), anti-GATA3 (Abcam, ab199428; 1:200), anti-TRIM37 (Abcam, ab95997; 1:200), and DAPI (nuclear marker). The expression of target proteins (red or green) and DAPI (blue) were examined by fluorescence microscopy; areas of co-localization are shown in the merged images.
For the mouse model, IF staining was performed on lung tissue sections derived from the indicated mouse models injected with Ad-K8-Cre that were fixed in 10% formalin (Fisher Scientific, Hampton, NH) and embedded in paraffin. Antigen retrieval (Citrate buffer pH 6.0, 20 min boil in microwave oven, level 1) was performed prior to blocking. Primary antibody [anti-KDM1/LSD1 (ab17721, 1:500, Abcam, Cambridge, UK)] and the secondary antibody [goat anti-rabbit IgG conjugated with AF594 (A11037, 1:250)] were used. Slides were counterstained with DAPI (1 μg/ml).
For the mouse model, IF staining was performed on lung tissue sections derived from the indicated mouse models injected with Ad-K8-Cre that were fixed in 10% formalin (Fisher Scientific, Hampton, NH) and embedded in paraffin. Antigen retrieval (Citrate buffer pH 6.0, 20 min boil in microwave oven, level 1) was performed prior to blocking. Primary antibody [anti-KDM1/LSD1 (ab17721, 1:500, Abcam, Cambridge, UK)] and the secondary antibody [goat anti-rabbit IgG conjugated with AF594 (A11037, 1:250)] were used. Slides were counterstained with DAPI (1 μg/ml).
3
Immunofluorescence Staining of Cancer Cells
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!