Western blots were generated by the semi-dry method. Protein lysates from cell lines were prepared using SIGMAFast protease inhibitor cocktail (Sigma). Proteins were transferred onto nitrocellulose membranes (Bio-Rad, München, Germany) and blocked with 5% dry milk powder dissolved in phosphate-buffered-saline buffer (PBS). The following antibodies were used: alpha-Tubulin (T6199, Sigma), HMX2 (NBP1-91997, Novus Biologicals, Abingdon, UK), HMX3 (LS-B8011, LSBio, Eching, Germany), ERK (sc-94, Santa Cruz Biotechnology, Heidelberg, Germany), phosphor (P)-ERK (sc-7383, Santa Cruz Biotechnology), EPX (LS-C354350, LSBio), HTR7 (LS-C358892, LSBio). For loading controls blots were reversibly stained with Poinceau (Sigma) and detection of alpha-Tubulin (TUBA) was performed thereafter. Secondary antibodies were linked to peroxidase for detection by Western-Lightning-ECL (Perkin Elmer, Waltham, MA, USA). Documentation was performed using the digital system ChemoStar Imager (INTAS, Göttingen, Germany).
Erk sc 94
Manufactured by Santa Cruz Biotechnology
Sourced in United States, Germany
ERK (sc-94) is a laboratory product offered by Santa Cruz Biotechnology. It is an antibody that recognizes the extracellular signal-regulated kinase (ERK) protein.
Automatically generated - may contain errors
Lab products found in correlation
Alpha tubulin, by Merck Group (2 mentions)
Western lightning ecl, by PerkinElmer (2 mentions)
Nitrocellulose membrane, by Bio-Rad (2 mentions)
Sigmafast protease inhibitor cocktail, by Merck Group (2 mentions)
Poinceau, by Merck Group (2 mentions)
Chemostar imager, by Intas (2 mentions)
E cadherin, by BD (1 mentions)
Vimentin, by BD (1 mentions)
Sb202190, by Merck Group (1 mentions)
Sp600125, by Merck Group (1 mentions)
P jnk, by Cell Signaling Technology (1 mentions)
Ab14106, by Abcam (1 mentions)
β actin, by Merck Group (1 mentions)
U0126, by InvivoGen (1 mentions)
P erk, by Santa Cruz Biotechnology (1 mentions)
P erk sc 7383, by Santa Cruz Biotechnology (1 mentions)
β actin sc 47778, by Santa Cruz Biotechnology (1 mentions)
β actin, by Sangon (1 mentions)
Amplex red acetylcholine acetylcholinesterase assay kit, by Thermo Fisher Scientific (1 mentions)
Ab15580, by Abcam (1 mentions)
5 protocols using erk sc 94
1
Western Blot Protein Detection Protocol
2
Oxidative Stress and Signaling in Cells
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Sodium trioxovanadate (NaVO3), oxovanadium sulfate (VOSO4), SB202190, SP600125, and 6-carboxy-2,7-dichlorodihydrofluorescein diacetate (DCFDA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). U0126 and MitoSOX Red fluorescence were from InvivoGen (San Diego, CA, USA) and Molecular Probes (Invitrogen, Carlsbad, CA, USA), respectively. Antibodies for E-cadherin (610182, BD) SMα (A5228, Sigma-Aldrich), SM22α (ab14106, Abcam, Cambridge, MA, USA), vimentin (550513, BD Biosciences, San Jose, CA, USA), β-actin (MAB1501, Millipore, MA, USA), IL-6 (ab6672, Abcam), p-p38 (#4511, CST), p38 (sc-7972, Santa Cruz, CA, USA), p-ERK (sc-7383, Santa Cruz), ERK (sc-94, Santa Cruz), p-JNK (#9251, Cell Signaling Danvers, MA, USA), JNK (#9258, Cell Signaling), p-p65 (Cell Signaling #3033), and p65 (#4764, Cell Signaling) were used in western blot analysis and immunohistochemistry.
3
Methamphetamine Effects on Neural Proteins
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Methamphetamine was obtained from Yunnan Province Public Security Department and dissolved in sterile water. Anti-mouse Trx-1 rabbit polyclonal antibody (14999-1-AP; 1:1,000) was purchased from ProteinTech (Wuhan, China). The antibodies MAG (sc-15324; 1:1,000), MBP (sc-376995; 1:1,000), procaspase-12 (sc-5627, 1:1,000), calpain1 (sc-13990; 1:1,000), phosphorylation extracellular signal-regulated kinase (p-ERK, sc-7383; 1:1,000), extracellular signal-regulated kinase (ERK, sc-94; 1:1,000), and β-actin (sc-47778; 1:1,000) were purchased from Santa Cruz Bio Technology (Santa Cruz, CA, USA). The antibodies procaspase-3 (19677-1-AP, 1:2,000) and nuclear factor κB (NF-κB) (10745-1-AP, 1:2,000) were purchased from Proteintech Group (Proteintech Group, Inc., Wuhan, Hubei, China). The antibody CDK5 (Ab-40773; 1:2,000) was purchased from Abcam (Abcam plc, Cambridge, UK). qRT-PCR primers [β-actin, interleukin-1beta (IL-1β), and tumor necrosis factor-alpha (TNF-α)] were purchased from Sangon Biotech Co., Ltd. (Shanghai, China).
4
Scopolamine and Piracetam Effects on Acetylcholine
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Scopolamine hydrobromide (P5295-25G) and piracetam (S1875-5G) were purchased from Sigma-Aldrich Corporation (St. Louis, MO, USA). The Amplex® Red Acetylcholine/Acetylcholinesterase Assay Kit (A12217) was purchased from Molecular Probes™ (Eugene, OR, USA). For western blot analysis, actin (A2066) was purchased from Sigma-Aldrich Corporation. ERK (sc-94) and Akt (sc-168) were supplied by Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against phosphorylated ERK (pERK, #9101), PI3K (#4257), phosphorylated PI3K (pPI3K, #4228), CREB (#9197), phosphorylated CREB (pCREB, #9196), and phosphorylated Akt (pAkt, #4058) were supplied by Cell Signaling Technology (Danvers, MA, USA). For immunohistochemical analysis, antibodies against pPI3K (ab61801, Y607), mature BDNF (mBDNF, ab75040), and Ki67 (ab15580) were supplied by Abcam (Cambridge, UK). The antibody against neuronal nuclei (NeuN, MAB377) was supplied by Millipore Corporation (Billerica, MA, USA), and that against pCREB (sc-7978) was supplied by Santa Cruz Biotechnology. The antibody against AchE (PA5-21371) was supplied by Thermo Fisher Scientific (Waltham, MA, USA). Fluorescein anti-rabbit (FI-1000), fluorescein anti-sheep (FI-6000), and Texas Red anti-mouse (TI-2000) antibodies were purchased from Vector Laboratories Inc. (Burlingame, CA, USA).
5
Western Blot Analysis of Cellular Proteins
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Western blots were generated by the semi-dry method. Protein lysates from cell lines were prepared using SIGMAFast protease inhibitor cocktail (Sigma, Taufkirchen, Germany). Proteins were transferred onto nitrocellulose membranes (Bio-Rad, München, Germany) and blocked with 5% dry milk powder dissolved in phosphate-buffered-saline buffer. The following antibodies were used: alpha-Tubulin (T6199, Sigma), VENTX (PA5-21006, Invitrogen, Darmstadt, Germany), ERK (sc-94, Santa Cruz Biotechnology, Heidelberg, Germany), and phospho(P)-ERK (sc-7383, Santa Cruz Biotechnology). For loading controls, blots were reversibly stained with Poinceau (Sigma) and detection of alpha-Tubulin (TUBA) performed thereafter. Secondary antibodies were linked to peroxidase for detection by Western-Lightning-ECL (Perkin Elmer, Waltham, MA, USA). Documentation employed the digital system ChemoStar Imager (INTAS, Göttingen, Germany).
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