Tumors and cells were lysed in RIPA lysis buffer with protease inhibitors (Roche) and phosphatase inhibitors (Sigma). Equal amount of total protein lysates were used for immunoblotting. The following antibodies were from Cell Signaling Technology (Beverly, MA) and were used at 1:1000 dilution: pAKT (#3787), AKT (#4691), pERK (#4370), ERK (#4695), pMET (#3077), MET (#3127), pEGFR (#3777), EGFR (#4267), pPRAS40 (#2997), PRAS40 (#2691), Vinculin (#13901). Other antibodies used in this paper are as follows: INPP4B (Abcam, Ab81269), beta-actin (Sigma, A2228).
Inpp4b
Manufactured by Abcam
Sourced in United States
INPP4B is an enzyme that catalyzes the conversion of phosphatidylinositol 4,5-bisphosphate (PIP2) to phosphatidylinositol 4-phosphate (PIP). It is involved in the regulation of cellular signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
Pras40, by Cell Signaling Technology (1 mentions)
P met, by Cell Signaling Technology (1 mentions)
Vinculin, by Cell Signaling Technology (1 mentions)
P egfr, by Cell Signaling Technology (1 mentions)
β actin, by Merck Group (1 mentions)
P pras40, by Cell Signaling Technology (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
P erk, by Cell Signaling Technology (1 mentions)
Phosphatase inhibitor, by Merck Group (1 mentions)
Protease inhibitor, by Roche (1 mentions)
Inpp4b, by Santa Cruz Biotechnology (1 mentions)
P akt ser473, by Cell Signaling Technology (1 mentions)
Total akt, by Cell Signaling Technology (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Pi 3 4 p2, by Echelon Biosciences (1 mentions)
Tau 1, by Merck Group (1 mentions)
Pi 3 4 p2, by Merck Group (1 mentions)
Alexa fluor 647, by Thermo Fisher Scientific (1 mentions)
Saponin, by Merck Group (1 mentions)
4 protocols using inpp4b
1
Immunoblotting for Signal Pathway Proteins
2
Molecular Biomarkers in Breast Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BCa cells were lysed for Western blotting and proteins were detected using the antibodies, GAPDH (Santa Cruz 32233), P-Ser473 AKT (Cell Signaling 4060), INPP4B (Santa Cruz 12318), Total AKT (Cell Signaling 9272), ERα (produced in our lab, SC1-1), and p27 (Santa Cruz 528). Tissue fixation and sectioning were processed as previously described [36 (link), 73 (link)]. Tissue sections were incubated with ERα antibodies (Santa Cruz, MC-20), P-Ser473 AKT (Cell Signaling 4060), INPP4B (Abcam EPR3108), Ki67 (Novocastra).
3
Immunofluorescence Staining of Neuronal Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were fixed at indicated time in 4% paraformaldehyde for 10 min and permeabilized with 0.1% Triton X-100 for 5 min at room temperature. For staining with anti-PI(3,4)P2 antibody, cells were permeabilized with 0.5% saponin (Sigma) for 15 min. After washing with PBS, the cells were blocked with 5% donkey serum in PBS for 1 h and incubated with primary antibody against GFP (Abcam, USA; 1:1000), Tau-1 (Millipore; 1:200), MAP2 (Millipore; 1:2000), INPP4B (Abcam; 1:50), and PI(3,4) P2 (Echelon, 1:200) overnight in 4°C. Appropriate secondary antibodies were incubated for 45 min at room temperature, and specimens were prepared as described in Immunohistochemistry. To label polymerized actin (F-actin), phalloidin-conjugated Alexa Fluor 647 (Invitrogen) were incubated together with secondary antibodies.
4
Standardized INPP4B Immunohistochemistry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We provided the tissue samples, and tissue microarray (TMA) manufacturing was done by Shanghai Biochip Company, Ltd., Shanghai, China. Briefly, the immunohistochemistry (IHC) process consisted of the following steps: baking, deparaffinization, antigen retrieval, quenching the endogenous peroxidase activity, blocking nonspecific staining, and incubation with the primary antibody (INPP4B, 1:50, Abcam) and the secondary antibody. The immunoreactivity score (IRS) was defined by “cell staining percentage×staining intensity”. The specimens were defined as negative (IRS =0~2) and positive (IRS=3~9). The detailed immunohistochemical process and staining evaluation were performed as previously described 20 (link).
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!