Mcp 1

Manufactured by Cell Signaling Technology

Sourced in United States

MCP-1 is a chemokine that binds to the CCR2 receptor and plays a role in the recruitment and activation of monocytes and macrophages. It is commonly used in cell biology research to study cellular processes related to inflammation and immune response.

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Close spelling variants of this product

Anti-MCP-1 (7 citations) Anti‐MCP‐1 antibody (2 citations) MCP-1 antibodies (2 citations)

27 protocols using mcp 1

Quantifying Monocyte Chemotactic Protein-1 in Ischemic Brain

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Protein lysates were obtained from dissection of ischemic (ipsilateral hemisphere) and healthy (contralateral hemisphere) tissues and subjected to SDS-PAGE prior to western blot analysis. Anti-mouse monocyte chemotactic protein-1 (MCP-1) (1:1000; Cell Signaling) and anti-mouse β-actin (1:40000; EMD Millipore) primary antibodies were used prior to detection by horseradish peroxidase (HRP)-conjugated secondary antibodies and revelation by enhanced chemiluminescence plus (ECL) solution (GE Healthcare Life Sciences). Blots were digitized and proteins level were densitometrically analyzed with ImageJ software.

Thériault P., Le Béhot A., ElAli A, & Rivest S. (2016). Sub-acute systemic erythropoietin administration reduces ischemic brain injury in an age-dependent manner. Oncotarget, 7(24), 35552-35561.

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Immunoblotting of Bovine Hepatocytes

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Immunoblotting was essentially conducted as previously described [30 (link)]. In brief, total protein was extracted from bovine hepatocytes using RIPA lysis buffer (Beyotime, Shanghai, China). The concentration of the proteins extracted was quantified with BCA methods (Pierce, Rockford, IL, USA). Equal amounts of protein were adjusted for separation on 10% SDS polyacrylamide gels. The separated proteins were transferred onto nitrocellulose membranes (Millipore, Billerica, MA, USA) and incubated with primary antibodies (Cell signaling technology) overnight at 4 °C on the rotator. The primary antibodies for p-p65, p65, p-IκB, IκB, IL-1β, p-JNK, JNK, p-ERK, ERK, p-p38, p38, p-c-Jun, c-Jun, ICAM-1, MCP-1, Nrf2, NQO1, HMOX1, and GAPDH were purchased from Cell Signaling Technology (#3033, #8242, #2859, #4812, #4668, #9252, #4370, #4695, #4511, #8690, #3270, #9165, #67,836, #81,559, #12,721, #62,262, #26,416, and #5174) and diluted as 1:1000. After moderate washing with TBST, the blots were incubated with horseradish peroxidase-coupled secondary antibodies (#7074, CST) and diluted as 1:5000. The intensity of each blot was normalized with quantification of GAPDH. The blots were analyzed and quantified with Image J software (LOCI).

Xu T., Liu R., Zhu H., Zhou Y., Pei T, & Yang Z. (2022). The Inhibition of LPS-Induced Oxidative Stress and Inflammatory Responses Is Associated with the Protective Effect of (-)-Epigallocatechin-3-Gallate on Bovine Hepatocytes and Murine Liver. Antioxidants, 11(5), 914.

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Investigating Neuroinflammatory Mechanisms

Cited 2 times

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All chemicals, including anti-CT19 amyloid antibody and 10% neutral buffered formalin solution used in this study, were from Sigma Chemical (St. Louis, MO, USA). Specific antibodies to MAPKs, phospho-MAPKs, total tau, p-tau-T181, p-tau-S396, β-amyloid, TNF-α, MCP-1, GFAP, cleaved caspase-3, or IL-6 were purchased from Cell Signaling (Danver, MA, USA). Specific antibody to GSK-3β, p-GSK-3β-S9, CDK5, p-tau-T231, iNOS, 3-NT, CYP2E1, or NeuN was from Abcam (Cambridge, MA, USA). Antibody against p-CDK5-Tyr15, cleaved caspase-3, Bax, GAPDH, NeuN, GFAP, HSP90, HNF-1α, BNIP3, and secondary antibodies conjugated with horse radish peroxidase were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Recombinant HIV-1 Tat and gp120 (B.MN D11 strain) proteins were provided by the NIAID AIDS Reagent Program (National Institutes of Health, Bethesda, MD, USA). Neuro-2a cells were purchased from the ATCC (American Type Culture Cells (Manassas, VA). Other materials not described here were the highest grades available and/or the same, as recently described [21 (link), 22 (link)].

Cho Y.E., Lee M.H, & Song B.J. (2017). Neuronal Cell Death and Degeneration through Increased Nitroxidative Stress and Tau Phosphorylation in HIV-1 Transgenic Rats. PLoS ONE, 12(1), e0169945.

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Protein Expression Profiling

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Primary antibodies used were CKS2 (Cat No. ab155078; Abcam), BMF (Cat No. 50542S; Cell Signaling), CD137 (34594S; Cell Signaling), MCP-1 (Cat No. 81559S; Cell Signaling), HSP90 (Cat No. 8165S; Cell Signaling). Proteins were visualized with chemiluminescent substrate (Cat No. 34577; Thermo Scientific) and a developing machine (Evolve).

Zhang M., Lotfollahzadeh S., Elzinad N., Yang X., Elsadawi M., Gower A., Belghasem M., Shazly T., Kolachalama V.B, & Chitalia V. (2023). Alleviating iatrogenic effects of paclitaxel via anti-inflammatory treatment. Research Square.

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Protein Expression Analysis of Inflammatory Markers

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Antibodies to PRR (anti-ATP6IP2/ab40790, Abcam, Cambridge, MA, USA), TNF-α (Santa Cruz biotechnology, Inc., Santa Cruz, CA, USA), COX-2 (Novus bilogicals, Colorado, USA), NF-κB p65 (Abcam, Cambridge, MA, USA) and MCP-1 (Cell Signaling, USA) were used in the Western blot (n=4) as previously described (12 (link)). Protein expression was normalized to β-actin protein.

Quadri S.S., Culver S, & Siragy H.M. (2017). PRORENIN RECEPTOR MEDIATES INFLAMMATION IN RENAL ISCHEMIA. Clinical and experimental pharmacology & physiology, 45(2), 133-139.

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Western Blot Analysis of Liver Proteins

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Liver lysates were resolved by SDS-PAGE and immunoblotted with specific antibodies (Chan et al., 2013 (link)). Antibodies were diluted 1:500 (for HSF1, HSP72 and HSP90) or 1:1000 (for other antibodies) with a TBST buffer containing 1% BSA, 0.02% sodium azide, and 0.0025% phenol red (Sigma-Aldrich, Australia). Antibodies for HSP72 and HSP90 were purchased from Enzo Life Sciences (Farmingdale, United States); MCP-1, HSF1, TGFβ, Smad3, mammalian target of rapamycin (mTOR), phospho^Ser2448 mTOR, α-Tubulin and GAPDH were purchased from Cell Signaling (Danvers, MA, United States). A nod-like receptor pyrin containing 3 (NLPR3) was purchased from AdipoGen (San Diego, United States). Goat Anti Mouse, Goat Anti Rabbit and Goat Anti-Rat from Santa Cruz (United States). Proteins were quantified using a ChemiDoc, and densitometry analysis was performed using Image Lab software (Bio-Rad Laboratories, Australia).

Mahzari A., Li S., Zhou X., Li D., Fouda S., Alhomrani M., Alzahrani W., Robinson S.R, & Ye J.M. (2019). Matrine Protects Against MCD-Induced Development of NASH via Upregulating HSP72 and Downregulating mTOR in a Manner Distinctive From Metformin. Frontiers in Pharmacology, 10, 405.

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Protein Expression and Signaling Analysis

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Antibodies for Akt, phospho-Akt (Ser⁴⁷³), AMP-activated protein kinase (AMPKα), phospho-AMPKα (Thr¹⁷²), the Akt substrate regulating GLUT4 translocation (AS160), phospho-AS160 (Thr⁶⁴²), ERK1/2, phospho-ERK1/2 (Thr²⁰²/Tyr²⁰⁴), p38 mitogen-activated protein kinase (p38), phospho-p38 (Thr¹⁸⁰/Tyr¹⁸²), IRS-1, IL-6, MCP-1, Na⁺/K⁺-ATPase, TNF-α, and α-Tubulin were purchased from Cell Signaling (Danvers, MA, USA). Ang II, IL-17A, COX-2, c-Jun-N-terminal kinase (JNK), phospho-JNK (Thr¹⁸³/Tyr¹⁸⁵), mineralocorticoid receptor (MCR), PPARɣ, C/EBPα, adipocyte protein (aP2), adiponectin, β-actin, α-adducin-1(ADD1), cytochrome P450 family 11-subfamily β-2 (CYP11β-2), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were purchased from Santa Cruz (Dallas, Texas, USA). Antibodies for leptin, mTOR, and SREBP-1c were obtained from Abcam (Cambridge, MA, USA). For rt-PCR, the primers of target cDNAs such as fatty acid synthase (FAS) and acetyl-CoA carboxylase (ACC) were purchased. (GenoTech Corp, Dajeon, Korea) ELISA kits for detecting the cytokines such as MCP-1, IL-6, and TNF-α were purchased from BioLegend Inc (San Diego, CA, USA). U0126 or PD98059, f MEK/ERK inhibitors were purchased from Sigma Aldrich (St. Louis, MO, USA)

Lee M., Sorn S.R., Lee Y, & Kang I. (2019). Salt Induces Adipogenesis/Lipogenesis and Inflammatory Adipocytokines Secretion in Adipocytes. International Journal of Molecular Sciences, 20(1), 160.

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Western Blot Analysis of Fatty Acid Transporters

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Liver (40 mg) or subcutaneous fat (100 mg) was homogenized using a Bullet Blender. Lysates from cultured MΦ (30 µg), liver homogenates (30 µg), or visceral fat homogenates (80 µg) were separated by gel electrophoresis and transferred onto nitrocellulose or PVDF membranes. Membranes were incubated overnight with an antibody against SLC27A4/FATP4 (ab200353, Abcam) or CEBPα (clone EP709Y, Cat. No. 1704-1, Epitomics). Primary antibodies purchased from Cell Signaling (Frankfurt, Germany) were antibodies against MCP-1 (Cat. No. 2029), PPARγ (Cat. No. 2435), p-FoxO1 (Ser256; Cat. No. 9461), ACC (Cat. No. 3662), FASN (C20G5, Cat. No. 3180), and GAPDH (14C10, Cat. No. 2118). After incubation with a secondary HRP-linked antibody, proteins were visualized by using Luminata Forte Western HRP Substrate (Millipore, Darmstadt, Germany). Quantification of proteins was carried out using ImageJ after normalization to GAPDH.

Göcebe D., Jansakun C., Zhang Y., Staffer S., Tuma-Kellner S., Altamura S., Muckenthaler M.U., Merle U., Herrmann T, & Chamulitrat W. (2023). Myeloid-specific fatty acid transport protein 4 deficiency induces a sex-dimorphic susceptibility for nonalcoholic steatohepatitis in mice fed a high-fat, high-cholesterol diet. American Journal of Physiology - Gastrointestinal and Liver Physiology, 324(5), G389-G403.

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Antibodies for Cell Signaling Pathways

Cited 1 time

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This has been performed as previously described [12 (link)]. Antibodies directed against Vimentin (RV202), Twist1, N-cadherin and interleukin-6 (IL-6) were purchased from Abcam (Cambridge, MA); Akt, phospho-Akt (193H12), Erk1/2, phospho-Erk1/2 (THR202/TYR204), E-cadherin (24E10) and MCP-1 from Cell Signaling (Danvers, MA); c-Myc from BD Biosciences (San Jose, CA); Cyclin D1 (HD11), ERα (F-10) and glyceraldehydes-3-phosphate dehydrogenase (GAPDH, FL-335), were purchased from Santa Cruz (Santa Cruz, CA).

Al-Howail H.A., Hakami H.A., Al-Otaibi B., Al-Mazrou A., Daghestani M.H., Al-Jammaz I., Al-Khalaf H.H, & Aboussekhra A. (2016). PAC down-regulates estrogen receptor alpha and suppresses epithelial-to-mesenchymal transition in breast cancer cells. BMC Cancer, 16, 540.

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Protein Expression Analysis in Tissue Lysates

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Tissue was lysed in radio-immuno-precipitation assay buffer (RIPA) (Boston BioProducts, Ashland, MA), 40ug were fractionated by sodium dodecyl sulfide polyacrylamide gel electrophoresis (SDS-PAGE) using 3%−8% Tris-Acetate gels (NuPage Novex Mini Gel), and the protein was transferred to polyvinylidene diflouride membranes (PVDF) (Millipore, Billerica, MA) and incubated overnight at 4 degrees C with primary antibodies against Transforming growth factor β (TGF-β), Janus family of tyrosine kinase-2 (Jak2), STAT 3, matrix metallopeptidase-9 (MMP-9), SMAD2/3, monocyte chemoattractant protein-1 (MCP-1), α-tubulin, N- Cadherin, α-fodrin, desmin, connexin 43, pan keratin, ß-tubulin, Troponin I, vimentin, filamin, ß-Actin and troponin T (Cell Signaling, Danvers, MA). The following day, membranes were incubated with the appropriate horseradish peroxidase (HRP)-linked secondary antibody for 1h at room temperature (Jackson ImmunoResearch, West Grove, PA). Immune complexes were visualized with enhanced chemi luminescence, images were captured with a digital camera system (G-Box, Syngene, Cambridge, England), and band densitometry was quantified as arbitrary light units using Image J Software (National Institutes of Health, Bethesda, MD). Anti-GAPDH antibody (Cell Signaling) was used on all membranes to correct for loading error. Representative images have been included in the manuscript.

Potz B.A., Sabe A.A., Sabe S.A., Lawandy I.J., Abid M.R., Clements R.T, & Sellke F.W. (2020). Calpain Inhibition Decreases Myocardial Fibrosis in Chronically Ischemic Hypercholesterolemic Swine. The Journal of thoracic and cardiovascular surgery, 163(1), e11-e27.

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