Total cells and tissues were prepared with RIPA buffer containing proteinase inhibitor (Pierce, Rockford, IL, USA). NE-PER™ Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher Scientific) were used to separate the cytosolic fraction and nuclear extracts. Supernatant samples were separated by 10% or 15% SDS-PAGE and transferred onto nitrocellulose membranes (Millipore, Billerica, WI, USA). The membranes were probed, after blocking with 5% nonfat milk, with primary antibodies (anti-PERK, Abcam, Ab229912; anti-eIF2, Abcam, Ab169528; anti p-eIF2, Abcam, Ab32157; anti-ATF4, Proteintech, 10835-1-AP; anti-CHOP, Proteintech, 15204-1-AP; anti-β-actin, Proteintech, 66009-1-lg; anti-NF-κB, Abcam, Ab16502; and anti-H3, Proteintech, 17168-1-AP) at 4°C overnight. After washing away the unbound antibody on the second day, the membranes were washed and incubated with HRP-conjugated rabbit secondary antibody (Beyotime) at room temperature for 1 h. The signals from the immunoreactive proteins were detected using the ECL reagent (Millipore).
Ab32157
Manufactured by Proteintech
Ab32157 is a laboratory equipment product. It is a tool used for a specific function in scientific research or analysis. No further details about its core function or intended use are provided, as an unbiased and factual description cannot be given without potentially extrapolating or interpreting beyond the available information.
Automatically generated - may contain errors
Lab products found in correlation
Ab2606, by Abcam (2 mentions)
Fluorescent secondary antibody, by LI COR (2 mentions)
Fluorescent total protein stain, by LI COR (2 mentions)
4e bp, by Abcam (2 mentions)
P 4e bp, by Abcam (2 mentions)
Ab 27792, by Santa Cruz Biotechnology (2 mentions)
Sc 11386, by Abcam (2 mentions)
P eif2α, by Abcam (2 mentions)
Eif2α, by Santa Cruz Biotechnology (2 mentions)
Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (2 mentions)
Nitrocellulose membrane, by Merck Group (1 mentions)
Ab229912, by Abcam (1 mentions)
Ab169528, by Abcam (1 mentions)
Hrp conjugated rabbit secondary antibody, by Beyotime (1 mentions)
Ecl reagent, by Merck Group (1 mentions)
3 protocols using ab32157
1
Western Blot Analysis of ER Stress Markers
2
Western Blot Analysis of SMA Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Western blot on tissues from SMA and control mice was performed exactly as described before (64 (link)). The following primary antibodies were used 4E-BP (Abcam; Ab-2606 (1:1000)); P-4E-BP (Abcam; Ab-27792 (1:1000)); eIF2α (Santa Cruz Biotechnology; Sc-11386 (1:1000)); p-eIF2α (Abcam; Ab32157 (1:1000)); Get4 (Proteintech 27768-1-AP (1:1000)); AE2 (acetylcholinesterase) (DSHB AE2DSHB (1:1000)). Antibody detection was performed using fluorescent secondary antibodies (LI-COR) and protein loading was normalized to a fluorescent total protein stain (LI-COR). Quantification was performed as described previously (34 (link)).
3
Western Blot Analysis of SMA Mice
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