Ab150113

Manufactured by Thermo Fisher Scientific

Ab150113 is a laboratory equipment product manufactured by Thermo Fisher Scientific. It serves as a core component in various scientific applications, providing essential functionality. The detailed specifications and intended uses of this product are not included in this response to maintain an unbiased and factual approach.

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Lab products found in correlation

4 protocols using ab150113

Engineered Human Skin Equivalents

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In all, 330,000 HDF per HSE were embedded in a collagen matrix containing rat tail collagen type I (Corning), 10× DMEM (Thermofisher) and sodium bicarbonate (Gibco/Invitrogen), filled into six-well-culture inserts (Thermofisher) and placed in deep six-well culture plates (Thermofisher). After 2 h of polymerisation at 37 °C, HSEs were equilibrated in DMEM medium supplemented with 10% foetal bovine serum (Sigma) and placed at 37 °C, 5% CO₂. After 3 days, 150,000 keratinocytes were seeded on top and submerged for 7 days in DMEM medium supplemented with 10% foetal bovine serum and growth factors (EGF, isoproterenol, hydrocortisone). Then the inserts were placed at the air–liquid interface for 7 days during which period the medium was supplemented with 1201.
HSEs were fixed in 10% formalin before embedding in paraffin and cutting into 5-µm-thick sections. Haematoxylin and eosin staining was performed using a standard protocol. Immunohistochemistry staining was performed using a standard protocol and the following primary antibodies: fillagrin (Abcam, ab17808), loricrin (Abcam, ab24722), keratin 10 (Abcam, ab9026) and the following secondary antibodies: α-mouse Alexa 488 (Abcam, ab150113) and α-rabbit Alexa 546 (Life technologies, A11010).

Lämmermann I., Terlecki-Zaniewicz L., Weinmüllner R., Schosserer M., Dellago H., de Matos Branco A.D., Autheried D., Sevcnikar B., Kleissl L., Berlin I., Morizot F., Lejeune F., Fuzzati N., Forestier S., Toribio A., Tromeur A., Weinberg L., Higareda Almaraz J.C., Scheideler M., Rietveld M., El Ghalbzouri A., Tschachler E., Gruber F, & Grillari J. (2018). Blocking negative effects of senescence in human skin fibroblasts with a plant extract. NPJ Aging and Mechanisms of Disease, 4, 4.

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Immunofluorescence Assay for Stem Cell Markers

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Immunofluorescence for Oct4 and Nestin was performed on adherent cells fixed with 4% paraformaldehyde. After fixation, cells were permeablized with 0.5% PBS/Triton for 10 min on ice. Following permeablization, cells were blocked in 10% FBS in 0.1% PBS/Tween for 1 hr at room temperature. After blocking, cells were incubated for 2 hr in primary antibody (Nestin: Millipore, MAB353, Oct4: Santa Cruz sc-8629), washed in 0.1% PBS/Tween and then incubated for 2 hr in secondary antibody (Goat anti-mouse 488 Life Technologies ab150113, Rabbit anti-goat 488 Life Technologies A27012). After washing, cells were stained with DAPI and imaged at 40x magnification.

Carter A.C., Xu J., Nakamoto M.Y., Wei Y., Zarnegar B.J., Shi Q., Broughton J.P., Ransom R.C., Salhotra A., Nagaraja S.D., Li R., Dou D.R., Yost K.E., Cho S.W., Mistry A., Longaker M.T., Khavari P.A., Batey R.T., Wuttke D.S, & Chang H.Y. (2020). Spen links RNA-mediated endogenous retrovirus silencing and X chromosome inactivation. eLife, 9, e54508.

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Immunofluorescence Staining of Pluripotency and Cardiac Markers

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Cells were fixed with 4% Paraformaldehyde (PFA) (Maokang Biotechnology, MM1504) for 15 min, permeabilized with 0.1% Triton X (Sangon Biotech, A110694) for 5 min, and blocked with 3% BSA (Sigma–Aldrich, A1933) for 1 h. Cells were subsequently stained with appropriate primary antibodies and AlexaFluor conjugated secondary antibodies including AlexaFluor® 647 (Abcam, ab150079, 1:500, RRID: AB_2722623), AlexaFluor® 594 (Abcam, ab150080, 1:500, RRID: ab150080; Abcam, ab150108, 1:500, RRID: AB_2732073) and AlexaFluor® 488 (Abcam, ab150113, 1:500, RRID: AB_2576208; Invitrogen, A11008, 1:500, RRID: AB_143165). Nuclei were stained with DAPI (Roche Diagnostics). For the staining of pluripotency markers, the primary antibodies were a goat anti-human OCT4 (Cell Signaling Technology, 2750S, 1:200, RRID: AB_823583), a rabbit anti-human NANOG (Santa Cruz Biotechnology, sc-33759, 1:200, RRID: AB_2150401), a mouse anti-human SSEA-4 (Abcam, ab16287, 1:200, RRID: AB_778073) and a rabbit anti-human SOX2 (Abcam, ab171380, 1:200, RRID: AB_2732072). For the staining of cardiac-specific markers, the primary antibodies were a mouse anti-human TNNT2 (Abcam, ab8295, 1:500, RRID: AB_306445) and a rabbit anti-human α-actinin (Cell Signaling Technology, 6487P, 1:100, RRID: AB_11179206). Pictures were taken with 60 × objective on confocal microscope (Nikon, A1) using NIS-Elements AR software (Nikon).

Sun Y., Su J., Wang X., Wang J., Guo F., Qiu H., Fan H., Cai D., Wang H., Lin M., Wang W., Feng Y., Fu G., Gong T., Liang P, & Jiang C. (2023). Patient-specific iPSC-derived cardiomyocytes reveal variable phenotypic severity of Brugada syndrome. eBioMedicine, 95, 104741.

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Immunofluorescence Staining of Stem Cell and Cardiomyocyte Markers

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Cells were fixed with 4% paraformaldehyde (PFA) for 15 min, permeabilized with 0.1% Triton X-100 (Sangon Biotech, A110694) for 5 min, and blocked with 3% bovine serum albumin (Sigma-Aldrich, A1933) for 1 h. Cells were subsequently stained with appropriate primary antibodies and AlexaFluor conjugated secondary antibodies. Primary antibodies include NANOG (Santa Cruz Biotechnology, sc-33759, 1:200), SOX2 (Abcam, ab97959, 1 μg/ml), OCT4 (Santa Cruz Biotechnology, sc-8628, 1:500), SSEA-4 (Abcam, ab16287, 1:500) TNNT2 (Abcam, ab45932, 1:400), α-actinin (Abcam, ab137346, 1:500), Na_v1.5 (Alomone labs, ASC-005, 1:200) and β-catenin (Abcam 237983, 1ug/ml). Secondary antibodies include AlexaFluor® 647 (Abcam, ab150079, 1:500), AlexaFluor® 594 (Abcam, ab150108, 1:500) and AlexaFluor® 488 (Abcam, ab150113, 1:500; Invitrogen, A11008, 1:500). Nuclei were stained with DAPI (Roche Diagnostics, 1023276001, 1 μg/ml). Pictures were taken with 60 × objective on confocal microscope (Nikon, A1) using NIS-Elements AR software (Nikon).

Cai D., Wang X., Sun Y., Fan H., Zhou J., Yang Z., Qiu H., Wang J., Su J., Gong T., Jiang C, & Liang P. (2023). Patient-specific iPSC-derived cardiomyocytes reveal aberrant activation of Wnt/β-catenin signaling in SCN5A-related Brugada syndrome. Stem Cell Research & Therapy, 14, 241.

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