Total protein was extracted from pancreatic cancer tissues and cell lines by RIPA buffer (Solarbio, Beijing, China) and separated by 10% SDS-PAGE before being transferred onto a PVDF membrane (Millipore, Bedford, MA, USA). The membrane was then blocked with 5% BSA and probed using the primary antibodies against ARHGAP4 (Abcam), p-mTOR (Cell Signaling Technology, Beverly, MA, USA), mTOR (Cell Signaling Technology), PKM2 (Cell Signaling Technology), HIF-1α (Abcam), and GAPDH (Cell Signaling Technology). The detection was performed using HRP-labeled secondary antibody (Beyotime Institute of Biotechnology). GAPDH levels are shown as loading controls.
Arhgap4
Manufactured by Abcam
Sourced in United States
ARHGAP4 is a Rho GTPase activating protein that negatively regulates Rho GTPase activity. It plays a role in the regulation of the actin cytoskeleton.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Hrp labeled secondary antibody, by Beyotime (1 mentions)
P mtor, by Cell Signaling Technology (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Hif 1α, by Abcam (1 mentions)
Ripa buffer, by Solarbio (1 mentions)
P akt, by Abcam (1 mentions)
Phospho s6 ribosomal protein ser240 244, by Cell Signaling Technology (1 mentions)
Enhanced ecl kit, by Thermo Fisher Scientific (1 mentions)
S6 ribosomal protein, by Cell Signaling Technology (1 mentions)
Ripa reagent, by Beyotime (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Bcl2 associated x (bax), by Abcam (1 mentions)
2 protocols using arhgap4
1
Protein Expression Analysis in Pancreatic Cancer
2
Protein Expression Analysis in Poor Ovarian Responders
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total proteins were extracted and purified from granulosa cells from poor ovarian responders or KGN cells using RIPA reagent (Beyotime Biotechnology). The concentration of purified proteins was analyzed by using BCA Protein Assay Kit (Thermo Fisher Scientific). For western blot, 20 µg of protein was subjected to SDS-polyacrylamide gel, followed by transferring to PVDF membrane (Millipore). The membrane was then subjected to blocking with 5% BSA, then incubation with primary antibodies against ARHGAP4 (1:1000), Bax (1:2000) from Abcam, AKT (1:2000), pAKT (Ser473) (1:1000), S6 Ribosomal Protein (1:2000), Phospho-S6 Ribosomal Protein (Ser240/244) (1:1000), Cleaved Caspase-3 (1:1000) from Cell Signaling Technology and Actin (1:10000) from Sigma-Aldrich, and finally incubation with AffiniPure-conjugated corresponding secondary antibody (Sigma-Aldrich) (1:1000–5000). The targeted protein in membrane was visualized using an enhanced ECL kit (Thermo Fisher Scientific). The expression level of Actin was considered as control.
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