Total proteins were extracted by RIPA. Proteins were separated by SDS-PAGE and electro-transferred onto PVDF membranes. The membranes were soaked in buffer containing 5% BSA for 1 h and then incubated with primary antibodies including anti‐p-mTOR (5536s, 1:1000, Cell Signaling Technology), anti‐mTOR (2983s, 1:1000, Cell Signaling Technology), anti‐p-PI3K (bs-6417R, 1:1000, Bioss), anti‐PI3K (bs-10657R, 1:1000, Bioss), anti‐p-AMPK (2531S, 1:1000, Cell Signaling Technology), anti‐AMPK (5832S, 1:1000, Cell Signaling Technology), anti‐p-AKT (Sc-293125, 1:1000, Santa), anti‐AKT (Sc-5298, 1:1000, Santa), anti‐PGC1α (bs-7535R, 1:1000, Bioss), anti‐LC3B (ab51520, 1:5000, Abcam), anti‐ZO-1 (61–7300, 1:1000, Invitrogen), anti‐Occludin (71–1500, 1:1000, Invitrogen), or anti‐Claudin-5 (34–1600, 1:1000, Invitrogen), at 4 °C overnight. Thereafter, the membranes were washed with Tris‐buffered saline Tween buffer and incubated with HRP‐rabbit (ab191866, 1:5000, Abcam) or HRP‐mouse (bs-0296G-HRP, 1:1000, Bioss) conjugated secondary antibody for 1 h at room temperature (RT). An Odyssey Infrared Imaging System was used to scan the membranes for further analysis. β-actin (CL594-66009, 1:1000, Proteintech) was used as loading control. Band intensities were quantified using Image J software.
Anti pi3k bs 10657r
Manufactured by Bioss Antibodies
Sourced in China
The Anti‐PI3K (bs-10657R) is a polyclonal antibody designed to detect phosphoinositide 3-kinase (PI3K), a key enzyme involved in various cellular processes. This antibody can be used for the detection and analysis of PI3K expression in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Ab51520, by Abcam (1 mentions)
Ab191866, by Abcam (1 mentions)
Anti p mtor 5536s, by Cell Signaling Technology (1 mentions)
Anti mtor 2983s, by Cell Signaling Technology (1 mentions)
Anti ampk 5832s, by Cell Signaling Technology (1 mentions)
Bs 0296g hrp, by Bioss Antibodies (1 mentions)
Anti occludin, by Thermo Fisher Scientific (1 mentions)
Anti claudin 5, by Thermo Fisher Scientific (1 mentions)
β actin, by Proteintech (1 mentions)
Gapdh, by ABclonal (1 mentions)
2 protocols using anti pi3k bs 10657r
1
Protein Expression Analysis by WB
2
Multifaceted Analysis of Protein Expression
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Total protein was extracted from mouse lung tissue and BEAS-2B cells, quantified, and diluted to the same concentration with 4× loading buffer. Proteins were separated on 10% or 8% SDS-PAGE gels and transferred to PVDF membranes. (Millipore, Billerica, MA, USA). After blocking with TBS-Tween 20 containing 5% skim milk for 3 h at room temperature, the membranes were incubated with primary antibodies for E-cadherin (Cat. No. 20874-1-AP), N-cadherin (Cat. No. 22018-1-AP), α-SMA (Cat. No. 14395-1-AP), AKT (Cat. No. 60203-2-lg), phospho-AKT (Cat. No. 66444-1-lg), HIF-1α (Cat. No. 20960-1-AP) and GAPDH (Cat. No. 10494-1-AP) were obtained from Proteintech (Wuhan, China), Anti-PI3K (bs-10657R) was ordered from Bioss (Beijing, China). Anti-p-PI3K (AP0854) was obtained from ABclonal (Wuhan, China) overnight at 4 °C with GAPDH as an internal control. The membrane was then washed with 1× TBST, incubated with the corresponding secondary antibody for 1 h, and washed again with 1× TBST. The density of the protein bands was analyzed using ImageJ software.
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