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P hsp27 s82

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P-Hsp27 (S82) is a primary antibody that recognizes the phosphorylated form of the heat shock protein Hsp27 at serine residue 82. It is used to detect and quantify the levels of this post-translationally modified protein in various biological samples.

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Lab products found in correlation

Ab17942, by Abcam (1 mentions) Ecl western blotting detection reagent, by GE Healthcare (1 mentions) Hsp27, by Cell Signaling Technology (1 mentions) P atf2, by Cell Signaling Technology (1 mentions) Eif4e, by Cell Signaling Technology (1 mentions) P eif4e, by Cell Signaling Technology (1 mentions) P p90rsk, by Cell Signaling Technology (1 mentions) P erk1 2 p p44 42, by Cell Signaling Technology (1 mentions) Phosstop and complete phosphatase protease inhibitor cocktails, by Roche (1 mentions) P mnk1, by Cell Signaling Technology (1 mentions) Jnk in 8, by Merck Group (1 mentions) Gsk 429286, by Merck Group (1 mentions) Cell culture supplies, by Thermo Fisher Scientific (1 mentions) Crt0066101, by Merck Group (1 mentions) Pakt s473 9271, by Cell Signaling Technology (1 mentions) P s6 s235 6, by Cell Signaling Technology (1 mentions)

2 protocols using p hsp27 s82

1

Western Blot Analysis of Cell Signaling Pathways

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Total protein
extracts were generated using lysis buffer (50 mM Tris-HCl, pH 7.4,
150 mM NaCl, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS, 1
mM EDTA) supplemented with PhosSTOP and Complete Phosphatase/Protease
Inhibitor Cocktails (Roche Diagnostics GmbH, Mannheim, Germany). Protein
extracts (20–25 μg per sample) were loaded onto sodium
dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels
and transferred electrophoretically to poly(vinylidene fluoride) (PVDF)
membranes. The following primary antibodies were used at a dilution
of 1:1000: ERK1/2 (p44/42) (ab17942, Abcam), p-ERK1/2 (p-p44/42) (Thr202/Tyr204)
(9101, Cell Signaling), MNK1 (2195, Cell Signaling), p-MNK1 (Thr197/202)
(2111, Cell Signaling), eIF4E (9742, Cell Signaling), p-eIF4E (p-Ser
209) (9741, Cell Signaling or NBP2-66802, Novus Biologicals), p38alpha
(9218, Cell Signaling), p38beta (2339, Cell Signaling), p-p38 (4511,
Cell Signaling), p-ATF2 (27934, Cell Signaling), Hsp27 (2402, Cell
Signaling), p-Hsp27 (S82) (2401, Cell Signaling), and p-p90 RSK (Thr573)
(9346, Cell Signaling). The primary HRP-conjugated antibody anti-β-actin
(Calbiochem) was used at a dilution of 1:20 000. Anti-mouse
and anti-rabbit HRP secondary antibodies were from Pierce and used
at a dilution of 1:10 000. Immunodetection of proteins was
performed using ECL Western Blotting Detection Reagents (GE Healthcare,
Buckinghamshire, U.K.).
Bou-Petit E., Hümmer S., Alarcon H., Slobodnyuk K., Cano-Galietero M., Fuentes P., Guijarro P.J., Muñoz M.J., Suarez-Cabrera L., Santamaria A., Estrada-Tejedor R., Borrell J.I, & Ramón y Cajal S. (2022). Overcoming Paradoxical Kinase Priming by a Novel MNK1 Inhibitor. Journal of Medicinal Chemistry, 65(8), 6070-6087.
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2

Endothelin-1 Signaling Pathway Analysis

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Chemicals were purchased from Sigma‐Aldrich unless otherwise stated. Cell culture supplies were purchased from Life Technologies. Endothelin 1 (CSCSSLMDKECVYFCHLDIIW) was purchased from Bachem (Product No. 4040254) and is referred to as EDN for the purpose of all stimulation experiments. The following key reagents were employed in the study: TiO2 (GL Sciences), 13C6‐15N2 lysine (Lys‐8, Silantes GmbH) and 13C6‐15N4 arginine (Arg‐10, Silantes GmbH). Heavy peptides for SRM were synthesised by JPT Peptide Technologies. Kinase inhibitors were purchased from Selleckchem, except for ddcAMP (Enzo Life Sciences), GSK‐429286, CRT0066101 and JNK‐IN‐8 (Sigma‐Aldrich). Primary antibodies and anti‐rabbit secondary antibodies (7074) were purchased from Cell Signalling Technologies [pAKT S473 (9271), pCREB S133 (9198), PKC motif (6967), PKA motif (9624), pS6 S235/6 (4858), pRB S807/11 (8516), CaMKII S286 (12716), pMAPKAPK2 T222 (3316), p‐cJun S73 (3270), p‐cRAF S338 (9427), pACC S79 (11818), pHSP27 S82 (9709), AKT (4691) and CREB (9197)]. Antibodies against β‐Actin (AC‐15), GAPDH (MAB374) and EDNRB (21,196) were purchased from Sigma‐Aldrich, Millipore and Santa Cruz, respectively. Anti‐mouse and anti‐goat secondary antibodies were purchased from Jackson ImmunoResearch.
Schäfer A., Gjerga E., Welford R.W., Renz I., Lehembre F., Groenen P.M., Saez‐Rodriguez J., Aebersold R, & Gstaiger M. (2019). Elucidating essential kinases of endothelin signalling by logic modelling of phosphoproteomics data. Molecular Systems Biology, 15(8), e8828.
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