MGF (purity ≥98%) was purchased from Chengdu Desite Biotechnology (Chengdu, China). β-estradiol (E8875), dimethyl sulfoxide (DMSO), and LPS were purchased from Sigma-Aldrich (St. Louis, MO, United States). Progesterone was obtained from VETEC (V900699). The antibodies used for western blotting were as follows: Iba1/AIF-1 (E4O4W) (#17198), GFAP (E4L7M) (#80788), PSD95 (D27E11) (#3450), BDNF (#47808), iNOS (D6B6S) (#13120), anti-p-ERK1/2 (Thr202/Tyr204) (#9101), anti-ERK1/2 (#9102), anti-p-p38 MAPK (Thr180/Tyr182) (#4511), anti-p38 MAPK (#9212), and anti-p-JNK (Thr183/Tyr185) (#9251) were purchased from Cell Signaling Technology (Beverly, MA, United States). β-tubulin (#CW0098A) and β-actin (#CW0096M) were procured from CWBiotech (Beijing, China).
Anti p p38 mapk thr180 tyr182
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-p-p38 MAPK-Thr180/Tyr182 is an antibody that detects phosphorylation of p38 MAPK at Thr180 and Tyr182. p38 MAPK is a serine/threonine protein kinase that plays a key role in the cellular response to various environmental stresses and inflammatory cytokines.
Automatically generated - may contain errors
Lab products found in correlation
Anti p38 mapk, by Cell Signaling Technology (4 mentions)
Anti p jnk thr183 tyr185, by Cell Signaling Technology (2 mentions)
Anti cdc2, by Cell Signaling Technology (2 mentions)
Anti p44 42 mapk erk1 2, by Cell Signaling Technology (2 mentions)
Anti akt, by Cell Signaling Technology (2 mentions)
Anti pakt s473, by Cell Signaling Technology (2 mentions)
Anti p p44 42 mapk erk1 2 thr202 tyr204, by Cell Signaling Technology (2 mentions)
Anti p cdc25c ser216, by Cell Signaling Technology (2 mentions)
Anti p histone h3 ser10, by Cell Signaling Technology (2 mentions)
Anti cdc25c, by Cell Signaling Technology (2 mentions)
Anti p cdc2 tyr15, by Cell Signaling Technology (2 mentions)
Brain derived neurotrophic factor (bdnf), by Cell Signaling Technology (1 mentions)
Anti p erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions)
Inos d6b6s, by Cell Signaling Technology (1 mentions)
β tubulin cw0098a, by CWBIO (1 mentions)
β actin, by CWBIO (1 mentions)
Anti erk1 2, by Cell Signaling Technology (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Anti p erk1 2, by Abcam (1 mentions)
Anti ppka, by Abcam (1 mentions)
8 protocols using anti p p38 mapk thr180 tyr182
1
Neuroinflammation Regulation by Molecules
2
Quantitative Immunoblotting for Signaling Proteins
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The following reagents and materials were used: anti-ALDH1B1 (ref. sc-374090) was purchased from Santa Cruz; anti-PI16 (ref. PAS 31882) from Thermo; anti-P70 S6K beta (ref: ab184551) from Abcam; and anti-Bcl-xL (ref. 2764), anti-pAkt (Ser473) (ref. 4060), anti-Akt (ref. 4685), anti-pMEK1/2 (Ser217/221) (ref. 9154), anti-MEK1/2 (ref. 9126), anti-pERK1/2 (Thr202/Tyr204) (ref. 4370), anti-ERK1/2 (ref. 9102), anti-pPKA (Thr197) (ref. 5661), anti-PKA (ref. 4782), anti-pSEK1/MKK4 (Ser257/Thr261) (ref. 9156), anti-SEK1/MKK4 (ref. 9152), anti pSAPK/JNK (Thr183/Tyr185) (ref. 9255), anti pSAPK/JNK (ref. 9252S), anti pMKK3-6 (Thr183/Tyr185) (ref. 9231), anti MKK3 (ref. 5674), anti p-p38 MAPK (Thr180/Tyr182) (ref. 4511), anti p38 MAPK (ref. 9212) were purchased from Cell Signaling. Electrophoresis reagents were purchased from Biorad and trypsin from Promega.
3
Protein Expression and Phosphorylation Analysis
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Cells were collected and lysed with NP-40 lysis buffer with protease inhibitors. Lysates were analyzed for protein expression/phosphorylation as described previously [25 (link),27 (link)]. The following antibodies were used at the indicated dilutions; all antibodies were purchased from Cell Signaling (Danvers, MA, USA): anti-p-AKT-S473 (1:1000, #4060), anti-AKT (1:1000, #4685), anti-p-p44/42 MAPK (Erk1/2) -Thr202/Tyr204 (1:1000, #4370), anti- p44/42 MAPK (Erk1/2) (1:1000, #4695), anti-p-p38 MAPK-Thr180/Tyr182 (1:1000, #9211), anti-p38 MAPK (1:1000, #8690), anti-p-cdc2-Tyr15 (1:1000, #4539), anti-cdc2 (1:1000, #9116), anti-p-CDC25C-Ser216 (1:1000, #4901), anti-cdc25C(1:1000, #4688), anti-p-Histone H3-ser10 (1:1000, #53348).
4
Quantifying Gene Expression in MDSCs
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Total RNA of MDSCs was extracted with the RNeasy kit (Qiagen, Valenica, CA, USA), and cDNA was synthesized using SuperScript III reverse transcriptase (Invitrogen, Carlsbad, CA, USA). An ABI 7900 real-time PCR system was used for quantitative PCR (qPCR), with primer and probe sets obtained from Applied Biosystems (Carlsbad, CA, USA). The results were analysed using SDS 2.1 software. The expression of each target gene is presented as the ‘fold change’ relative to that of control samples, as described previously.20 (link) Immunoblot analysis was performed as described previously.21 (link) Western blots were performed using specific antibodies: anti-GR (EPR4595; Abcam), anti-p-ErK (Thr202/Tyr204; Cell Signaling Technology, Danfoss, MA, USA), anti-p-JNK (Thr183/Tyr185; Cell Signaling Technology), anti-p-p38MAPK (Thr180/Tyr182; Cell Signaling Technology), anti-β-actin (AC-15; Sigma-Aldrich, St Louis, MO, USA) and anti-HIF1α (IC1935P; R&D system).
5
Protein Extraction and Immunoblotting Protocol
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Preparation of whole cell lysates and kidney tissue homogenates, and immunoblotting were performed as previously described [32 (link)]. The primary antibodies were obtained as follows: anti-α-SMA (ab5694; Abcam), anti-FN (ab2413; Abcam), anti-col-I (ab138492; Abcam), and anti-E-cadherin (sc-7870; Santa Cruz Biotechnology or ab15148; Abcam). Anti-p-Smad2 (Ser465/467)/Smad3 (Ser423/425) (8828), anti-p-Erk1/2 (Thr202/Tyr204) (9101), and anti-p-p38MAPK (Thr180/Tyr182) (4511) were obtained from Cell Signaling Technology (Danvers, MA, USA).
6
Antibody Panel for Cell Signaling
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Primary antibodies used were anti–Lamin A/C (catalog 2032), rabbit monoclonal anti–p-p38 MAPK (Thr180/Tyr182) (catalog 9215), anti–p-SAPK/JNK (Thr183/Tyr185) (catalog 9255) (all Cell Signaling Technology); anti–β-actin (catalog A5441) and anti–p-PPARγ S112 (catalog 04-816-I) (both Sigma-Aldrich); anti–p-ERK (sc-7383, Santa Cruz Biotechnology); anti-PPARγ (A0270, Abclonal); and anti-PPARα (catalog PAa-822A), anti-PPARδ (PA1-823A), and anti–p-PPARγ S273 (catalog BS-4888R) (all Thermo Fisher Scientific). See complete unedited blots in the supplemental material.
7
NGF Signaling Pathway Analysis
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Human recombinant NGF was kindly provided by Dr L. Manni (Institute of Translational Pharmacology, CNR, Rome, Italy). Antibodies (Abs) against β-actin, TrkA, p75, GFAP, p‐16, used for Western Blot were from Santa Cruz Biotechnology (Dallas, TX, United States). Abs anti‐extracellular signal regulated kinase (ERK)1/2, anti‐p‐ERK1/2 (Thr202/Tyr204), anti-AKT, anti‐p‐AKT (Ser473), anti-p38 MAPK and anti-pp-38 MAPK (Thr180/Tyr182) were from Cell Signaling Technology (Danvers, MA, United States); anti-MASH-1 Ab was from Thermo-Fisher (Waltham, MA, United States). Abs for immunofluorescence studies were against: GFAP (Thermo-Fisher), vimentin (Santa Cruz Biotechnology), synaptophysin, S100 β, nestin, all from Sigma-Aldrich. Alexa Fluor 488 F (Ab)2 fragment of goat anti-rabbit IgG and Alexa Fluor 488 F (ab)2 fragment of goat anti-mouse IgG were used as secondary Abs for IF (Jackson Immunoresearch, Ltd. Europe). Eukitt from Sigma-Aldrich was employed as mounting solution.
8
Western Blot Analysis of Cell Signaling
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Cells were collected and lysed with NP-40 lysis buffer with protease inhibitors. Lysates were analyzed for protein expression/phosphorylation as described previously [18, 19] . The following antibodies were used at the indicated dilutions; all antibodies were purchased from Cell Signaling: anti-p-AKT-S473 (1:1000, #4060), anti-AKT (1:1000, #4685), anti-p-p44/42 MAPK (Erk1/2) -Thr202/Tyr204 (1:1000, #4370), anti-p44/42 MAPK (Erk1/2) (1:1000, #4695), anti-p-p38 MAPK-Thr180/Tyr182 (1:1000, #9211), anti-p38 MAPK (1:1000, #8690), anti-p-cdc2-Tyr15 (1:1000, #4539), anti-cdc2 (1:1000, #9116), anti-p-CDC25C-Ser216 (1:1000, #4901), anti-cdc25C(1:1000, #4688), anti-p-Histone H3-ser10 (1:1000, #53348).
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