Ebio7979

Manufactured by Thermo Fisher Scientific

Sourced in United States

The EBio7979 is a laboratory instrument designed for electrophoresis. It provides a controlled environment for the separation and analysis of biomolecules, such as DNA, RNA, or proteins, based on their size and charge.

Automatically generated - may contain errors

Lab products found in correlation

Pvdf membrane, by Merck Group (2 mentions) A0398, by Agilent Technologies (1 mentions) A0452, by Agilent Technologies (1 mentions) Mab1004, by Merck Group (1 mentions) Mab1044, by Merck Group (1 mentions) E0413, by Agilent Technologies (1 mentions) Ab1839, by Merck Group (1 mentions) E0353, by Agilent Technologies (1 mentions) Sb203580, by Enzo Life Sciences (1 mentions) 5z 7 oxozeaenol, by Bio-Techne (1 mentions) Rapamycin, by Enzo Life Sciences (1 mentions) Sp600125, by Enzo Life Sciences (1 mentions) Phos tag acrylamide, by Fujifilm (1 mentions) Okadaic acid, by Enzo Life Sciences (1 mentions) Mouse anti flag, by Merck Group (1 mentions) λ ppase, by New England Biolabs (1 mentions) U0126, by Enzo Life Sciences (1 mentions) β actin 1 19, by Santa Cruz Biotechnology (1 mentions) Sds page gel, by Bio-Rad (1 mentions) Laemmli sample buffer, by Bio-Rad (1 mentions)

6 protocols using ebio7979

Immune Markers in Tissue Samples

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The following antibodies were used: polyclonal rabbit antibody against human myeloperoxidase (MPO) (A0398, Dakocytomation, Glostrup, Denmark) and cluster of differentiation 3 (CD3 (A0452, Dakocytomation, Glostrup, Denmark), monoclonal rabbit antibody against human fork head box P3 (FoxP3) (clone eBio7979, 14-7979-82, eBioscience, San Diego, CA, USA), ovine interleukin-6 (IL-6) (MAB1004, Millipore, Darmstadt, Germany), and ovine IL-8 (MAB1044 Millipore, Darmstadt, Germany). Antibodies against intestinal fatty acid binding protein (I-FABP) were kindly provided by the Department of Surgery, Maastricht University Medical Centre, the Netherlands. Secondary antibodies were the following: biotin-conjugated rabbit antimouse (E0413, DakoCytomation, Glostrup, Denmark), swine antirabbit (E0353, DakoCytomation, Glostrup, Denmark), and peroxidase-conjugated goat antirabbit (111-035-045, Jackson, West Grove, PA, USA). Detection antibodies against IL-6 (AB1839, Millipore, Darmstadt, Germany) and IL-8 (AB1840, Millipore, Darmstadt, Germany) were used.

van Gorp C., de Lange I.H., Spiller O.B., Dewez F., Cillero Pastor B., Heeren R.M., Kessels L., Kloosterboer N., van Gemert W.G., Beeton M.L., Stock S.J., Jobe A.H., Payne M.S., Kemp M.W., Zimmermann L.J., Kramer B.W., Plat J, & Wolfs T.G. (2019). Protection of the Ovine Fetal Gut against Ureaplasma-Induced Chorioamnionitis: A Potential Role for Plant Sterols. Nutrients, 11(5), 968.

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Investigating Foxp3 Regulation by NLK

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The following antibodies were used: rat anti-Foxp3 clone PCH101 and mouse anti-Foxp3 clone eBio7979 (eBioscience), mouse anti-Flag (Sigma-Aldrich), mouse anti-hemaglutinin (HA) clone 12CAS, rabbit anti-NLK (H-100) and goat anti-Actin (I-19) (Santa Cruz Biotechnology), and anti-HSP90 was purchased from Professor Ineke Braakman (UMC Utrecht, Utrecht, the Netherlands).
The following reagents were used: Phos-tag™ Acrylamide (WAKO Chemicals GmbH), Okadaic acid, SB203580">SB203580, SP600125">SP600125, U-0126 and Rapamycin (Enzo Lifesciences), PKB inhibitor VIII (Calbiochem), 5Z-7-Oxozeaenol and BIO (Tocris Bioscience, Bristol, United Kingdom, λPPAse (New England BioLabs) and recombinant GST-NLK active protein (Sigma-Aldrich).

Fleskens V., Minutti C.M., Wu X., Wei P., Pals C.E., McCrae J., Hemmers S., Groenewold V., Vos H.J., Rudensky A., Pan F., Li H., Zaiss D.M, & Coffer P.J. (2019). Nemo-like Kinase Drives Foxp3 Stability and Is Critical for Maintenance of Immune Tolerance by Regulatory T Cells. Cell Reports, 26(13), 3600-3612.e6.

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Immunoblotting Analysis of Cell Signaling

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Cells were lysed in RIPA lysis buffer containing protease inhibitor cocktail and phosSTOP (Roche). Cell lysate was treated with 2 × Laemmli sample buffer (Bio-Rad) and denatured at 100°C for 10 min. Protein extracts were separated by 4–15% SDS-PAGE gel (Bio-Rad) and transferred to the PVDF membrane (Millipore) and analyzed by immuno-blotting with following antibodies: β-actin (I-19, Santa Cruz), CDK8 (P455, CST), pStat1-Y701 (58D6, CST), pStat1-S727 (D3B7, CST), pSmad2 linker (Ser245/250/255, CST), pSmad2 (Ser465/467)/Smad3 (Ser425/427) (D27F4, CST), and FoxP3 (eBio7979, eBioscience). Images of western blot were taken by Bio-Rad imaging system and had been cropped according to the molecular weight for presentation.

Guo Z., Wang G., Lv Y., Wan Y.Y, & Zheng J. (2019). Inhibition of Cdk8/Cdk19 Activity Promotes Treg Cell Differentiation and Suppresses Autoimmune Diseases. Frontiers in Immunology, 10, 1988.

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Immunoblotting Analysis of Transcription Factors

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Sorted CD4⁺GFP⁻, CD4⁺GFP⁺, or CD4⁺CD25⁻ and CD4⁺CD25⁺ cells were lysed directly in SDS-loading buffer and resolved on 12% polyacrylamide gel (5×10⁴ cells/lane) and electrotransferred onto a PVDF membrane (Millipore). Membranes were probed with antibody specific for Foxp3 (1 μg/ml, eBio7979, eBioscience) as described (27 (link)). For Smad2 and phospho-Smad2 detection membranes were probed overnight (4°C) with the corresponding antibodies (Cell Signaling Technology). Images were scanned and analyzed using ImageQuant 5.2 software (GE).

Kuczma M., Wang C.Y., Ignatowicz L., Gourdie R, & Kraj P. (2015). Altered connexin 43 expression underlies age dependent decrease of Treg cell suppressor function in NOD mice. Journal of immunology (Baltimore, Md. : 1950), 194(11), 5261-5271.

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FoxP3 Western Blot Analysis

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Western blots of FoxP3 were performed as described before^{27 (link)}; intensity of FoxP3 bands detected with anti-FoxP3 antibody (eBio7979, eBioscience) was analyzed using ImageJ 1.48 v (NIH) software and values are calculated relative to the corresponding intensity of heat shock protein 70 (HSP70) bands detected with anti-HSP70 antibody (1B5, Assay designs).

Mailer R.K., Gisterå A., Polyzos K.A., Ketelhuth D.F, & Hansson G.K. (2017). Hypercholesterolemia Enhances T Cell Receptor Signaling and Increases the Regulatory T Cell Population. Scientific Reports, 7, 15655.

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FOXP3 Isoform Analysis by Western Blot

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Cells were lysed in 1% NP40 buffer. Protein lysates were loaded on an SDS gel electrophoresis (Bio-Rad) to analyze FOXP3 isoforms. Proteins were blotted on Immobilon P membranes (Millipore) using a Transblot (Bio-Rad) and detected with anti-FOXP3total (eBio7979, eBioscience). After incubation of the membrane with the HRP-coupled anti-mouse secondary antibody (Santa Cruz) and ECL Western Blot detection reagent (Amersham), chemoluminescence was detected on a ChemiDoc MP Imaging System (Bio-Rad).

Joly A.L., Seitz C., Liu S., Kuznetsov N.V., Gertow K., Westerberg L.S., Paulsson-Berne G., Hansson G.K, & Andersson J. (2018). Alternative Splicing of FOXP3 Controls Regulatory T Cell Effector Functions and Is Associated With Human Atherosclerotic Plaque Stability. Circulation research, 122(10).

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