The epidermis, midgut, and fat body of the 6th-48 h and 6th-96 h larvae were extracted using lysate buffer (P0013J, Beyotime Biotechnology, Shanghai, China) with 1 mM phenylmethanesulfonyl fluoride (PMSF; A100754, Sangon Biotech), followed by centrifugation at 10000 × g at 4°C for 10 min. The supernatant was incubated with λPPase according to the kit’s instruction (P0753L, New England Biolabs, Ipswich, MA, USA), and analyzed using western blotting.
A100754
Manufactured by Sangon
Sourced in China
The A100754 is a laboratory equipment product. It is designed to perform a core function within the laboratory environment. The detailed specifications and intended use of this product are not available.
Automatically generated - may contain errors
Lab products found in correlation
Ab15050, by Abcam (1 mentions)
T perk, by Cell Signaling Technology (1 mentions)
Xbp1s e9v3e, by Cell Signaling Technology (1 mentions)
P perk, by Cell Signaling Technology (1 mentions)
Bip c50b12, by Cell Signaling Technology (1 mentions)
C ebpβ, by Abcam (1 mentions)
C ebpβ, by Cell Signaling Technology (1 mentions)
P eif2α, by Cell Signaling Technology (1 mentions)
Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
2 protocols using a100754
1
Protein Phosphatase Assay in Insect Larvae
2
Immunoblotting Analysis of ER Stress Markers
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For the IB assay, 1 × 104 LSK cells or 2 × 105 splenocytes were washed once with RPMI with 0.1% phenylmethylsulfonyl fluoride (A100754; Sangon Biotech), and the supernatant was carefully removed by using a 200-μl tip. The cell pellet was lysed in 10 μl of lysis buffer (62.5 mM Tris-HCl, pH 6.8, 2% [wt/vol] SDS, 10% [wt/vol] glycerol, 50 mM dithiothreitol, and 0.01% [wt/vol] bromophenol blue in ultrapure water) on ice. Equal amounts of cellular proteins were separated on 10% SDS-PAGE gels, immunoblotted with the indicated antibodies, and visualized using the SuperSignal West Pico Chemiluminescent Substrate (34580; Thermo Fisher Scientific). The primary antibodies used for IB were as follows: β-actin (AC-15; BM5422; Boster Biological Technology); BIP (C50B12; 3177, RRID: AB_2119845; Cell Signaling Technology); t-PERK (C33E10; 3192, RRID: AB_2095847; Cell Signaling Technology); p-PERK (16F8; 3179, RRID: AB_2095853; Cell Signaling Technology); p-eIF2α (119A11; 3597, RRID: AB_390740; Cell Signaling Technology); ATF4 (D4B8; 11815, RRID: AB_2616025; Cell Signaling Technology); XBP1-S (E9V3E; 40435, RRID: AB_2891025; Cell Signaling Technology); C/EBPβ (#3087; 3087, RRID: AB_2078052; Cell Signaling Technology) for LAP; C/EBPβ (1H7; ab15050, RRID: AB_301598; Abcam) for LAP*/LAP/LIP; and CHOP (L63F7; 2895, RRID: AB_2089254; Cell Signaling Technology).
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