Cytomembrane protein and total protein were extracted from jejunum, ileum, and IPEC-J2 cells, using a membrane and cytosol protein extraction kit (Beyotime) and RIPA Lysis Buffer (Beyotime) in accordance with the manufacturer's instructions. Cytomembrane protein was used to determine the relative protein levels of AQP3 (sc-20811, Santa Cruz, TX, USA), AQP4 (ab46182, Abcam, LON, UK), AQP8 (sc-14984, Santa Cruz), epithelial sodium channel alpha subunit (α-ENaC, ab65710, Abcam), sodium-hydrogen exchanger 3 (NHE3, sc-28757, Santa Cruz), Cl−/HCO3− exchanger (DRA/PAT1, sc-161150, Santa Cruz), β-ENaC (14134-1-AP, Proteintech, CA, USA), and β-ATPase Na+/K+ antibody (ab254025, Abcam). Total protein was used to measure the relative protein levels of zonula occludens-1 (ZO-1, 21773-1-AP, Proteintech), occludin (ab31721, Abcam), E-cadherin (Proteintech), claudin-1 (#13995, Cell Signaling Technology, Boston, USA), claudin-2 (ab53032, Abcam), AMPKα1 (10929-2-AP, Proteintech), AMPKα2 (18167-1-AP, Proteintech), PGC-1α (ab106814, Abcam), target of rapamycin complex 2 (TORC2, 12497-1-AP, Proteintech), silent information regulator T1 (SIRT1,13161-1-AP, Proteintech), and β-actin (60008-1-Ig, Proteintech). These proteins were detected by the Western blotting technique as previously reported (Tan et al., 2010 (link)).
Ab254025
Manufactured by Abcam
Sourced in United States
Ab254025 is a lab equipment product offered by Abcam. It is a piece of lab equipment designed for a specific function within the scientific research process. The core function of this product is to perform a particular task or procedure in a laboratory setting, however, a detailed description of its intended use or capabilities is not available at this time.
Automatically generated - may contain errors
Lab products found in correlation
Ab31721, by Abcam (1 mentions)
Ab106814, by Abcam (1 mentions)
Ab46182, by Abcam (1 mentions)
Ab53032, by Abcam (1 mentions)
E cadherin, by Proteintech (1 mentions)
Membrane and cytosol protein extraction kit, by Beyotime (1 mentions)
Sirt1 13161 1 ap, by Proteintech (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Mycoplasma detection kit, by Vazyme (1 mentions)
Ab9100, by Abcam (1 mentions)
Flag tag (flag), by Merck Group (1 mentions)
Ccnd1, by Cell Signaling Technology (1 mentions)
Anti rabbit igg, by Cell Signaling Technology (1 mentions)
γh2ax, by Cell Signaling Technology (1 mentions)
Cleaved parp 1, by Cell Signaling Technology (1 mentions)
Hrp linked anti mouse igg, by Cell Signaling Technology (1 mentions)
α tubulin, by Cell Signaling Technology (1 mentions)
C myc, by Cell Signaling Technology (1 mentions)
Parp1, by Cell Signaling Technology (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
2 protocols using ab254025
1
Intestinal Membrane Protein Analysis
2
Characterization of Human Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human NPC cell lines (S26 and 5–8 F) were kindly gifted by Professor Chaonan Qian at SYSUCC (Guangzhou, China). The human embryonic kidney HEK293T cells were obtained from Cell Bank of Type Culture Collection of Chinese Academy of Sciences, Chinese Academy of Sciences. All cells were maintained in DMEM (Dulbecco’s Modified Eagle Medium, Gibco, NY, USA) with 10% fetal bovine serum (FBS, Gibco, NY, USA) and 1% Penicillin-Streptomycin (100 nM Gibco, NY, USA), in the humidified incubator with 5% CO2 at 37 °C. All cell lines were routinely examined to be mycoplasma-free using the mycoplasma detection kit (Vazyme, Nanjing, China).
Primary antibodies were commercially available: PARP1, Cleaved-PARP1, CCND1, C-MYC, N-Cadherin, Vimentin, γ-H2AX, α-Tubulin (9542, 5625, 2922, 9402, 14215, 5741, 9718, 2144, Cell Signaling Technology, Danvers, USA), Ki-67 (AB_393778, BD Pharmingen™, New Jersey, USA), SRSF3, HA, PMCA, EZH2 (ab125124, ab9100, ab254025, ab191250, Abcam, Cambridge, USA), AMOTL1, YAP1 (16871-1-AP, 16871-1-AP13584-1-AP, Proteintech, Chicago, USA), GAPDH (RM2002, Ray Antibody Biotech, Beijing, China), β-actin (AC004, Abclonal, Wuhan, China), Flag-tag (F1804, Sigma-Aldrich, St. Louis, USA). Secondary antibodies were HRP-linked anti-mouse IgG and anti-rabbit IgG (7076, 7074, Cell Signaling Technology, Danvers, USA).
Primary antibodies were commercially available: PARP1, Cleaved-PARP1, CCND1, C-MYC, N-Cadherin, Vimentin, γ-H2AX, α-Tubulin (9542, 5625, 2922, 9402, 14215, 5741, 9718, 2144, Cell Signaling Technology, Danvers, USA), Ki-67 (AB_393778, BD Pharmingen™, New Jersey, USA), SRSF3, HA, PMCA, EZH2 (ab125124, ab9100, ab254025, ab191250, Abcam, Cambridge, USA), AMOTL1, YAP1 (16871-1-AP, 16871-1-AP13584-1-AP, Proteintech, Chicago, USA), GAPDH (RM2002, Ray Antibody Biotech, Beijing, China), β-actin (AC004, Abclonal, Wuhan, China), Flag-tag (F1804, Sigma-Aldrich, St. Louis, USA). Secondary antibodies were HRP-linked anti-mouse IgG and anti-rabbit IgG (7076, 7074, Cell Signaling Technology, Danvers, USA).
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!