Mouse igg1

Manufactured by Santa Cruz Biotechnology

Sourced in United Kingdom, United States

Mouse IgG1 is an immunoglobulin subclass found in mice. It is a commonly used isotype control in flow cytometry and other immunoassays.

Automatically generated - may contain errors

Lab products found in correlation

Rabbit igg1, by Abcam (1 mentions) Sc 8059, by Santa Cruz Biotechnology (1 mentions) Rhe selectin, by R&D Systems (1 mentions) Anti lfa 1 clone ts2 4, by BioLegend (1 mentions) Rhp selectin, by R&D Systems (1 mentions) Rhil 8, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Penicillin, by Merck Group (1 mentions) Streptomycin, by Merck Group (1 mentions) Rat igg2b, by Thermo Fisher Scientific (1 mentions) Troponin t, by Thermo Fisher Scientific (1 mentions) Mouse igg1k, by BD (1 mentions) Cd172a b pe, by BioLegend (1 mentions) Cd11b, by Bio-Rad (1 mentions) Inside stain kit, by Miltenyi Biotec (1 mentions) Cd105, by BD (1 mentions) Cd166, by BD (1 mentions) Rabbit anti il 17a, by Abcam (1 mentions) Alexa fluor 594, by Santa Cruz Biotechnology (1 mentions) Ezview red protein a affinity gel, by Merck Group (1 mentions)

8 protocols using mouse igg1

Immunohistochemical Analysis of DAB2IP and p-STAT3

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IHC was performed on 3 μm-thick sections using standard protocols and an automated IHC staining system (Dako Autostainer Plus; Dako Denmark A/S, Glostrup, Denmark) as described previously.7 (link) The primary antibodies used were those targeting human DAB2IP (clone ab87811, diluted 1:400, rabbit IgG1; Abcam, Cambridge, UK) and p-STAT3 (sc-8059, diluted 1:500, mouse IgG1, Santa Cruz Biotechnology Inc., Dallas, TX, USA). Antigen retrieval was achieved using EDTA buffer (pH 9.0) in a pressure cooker for 4 min for DAB2IP or 20 min for p-STAT3. The stained tissue sections were reviewed and scored separately by two pathologists blinded to the clinical parameters. Discordant cases were discussed using a double-headed microscope to obtain a consensus classification.
The total immunostaining score for DAB2IP and p-STAT3 was calculated as the sum of the percentage of stained cells and the staining intensity, according to previously published reports.7 (link),11 (link) The percentage of stained cells was scored as 0 (0%), 1 (1%–25%), 2 (26%–50%), 3 (51%–75%), or 4 (>75%). The staining intensity was scored as 0 (no staining), 1 (weak staining), 2 (moderate staining), or 3 (strong staining). The staining of DAB2IP and p-STAT3 was determined as negative (low expression, a final staining score of <3) or positive (high expression, a final staining score of ≥3).

Zheng L., Chen K., Zhu L., Su D, & Cheng G. (2017). Low expression of DAB2IP predicts an unfavorable prognosis in human bladder carcinoma. OncoTargets and therapy, 10, 5719-5726.

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E-selectin and Integrin Adhesion Assay

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Rectangular glass capillaries were precoated with 500 µg/ml protein G for 2 h. Capillaries were washed with PBS and coated with 6.6 µg/ml recombinant human E-selectin (rhE-selectin; R&D Systems) and 25 µg/ml mouse IgG1 (Santa Cruz Biotechnology, Inc.), 25 µg/ml activation-specific anti–β₂ integrin (clone KIM127; ATCC), or 10 µg/ml anti–LFA-1 (clone TS2/4; BioLegend) for 1 h and subsequently blocked with 1% casein. In another set of experiments, capillaries were coated with 20 µg/ml rhP-selectin (R&D Systems), 50 µg/ml rhIL-8 (PeproTech), and 5 µg/ml mouse IgG1, 5 µg/ml activation-specific anti-β₂ integrin (clone mAb24; provided by N. Hogg, Cancer Research UK London Research Institute, London, England, UK), 10 µg/ml anti–LFA-1 (clone TS2/4), 40 µg/ml activation-specific anti–Mac-1 (clone CBRM1/5; BioLegend), or 30 µg/ml anti–Mac-1 (clone M1/70; BioLegend). HL60 cells were resuspended in freshly isolated human plasma. The flow chamber was perfused with the cell suspension for 2 min and subsequently flushed with PBS containing 1 mM MgCl₂ and CaCl₂ for 1 min to remove nonadherent cells. 10 representative fields of view were recorded, and the number of adherent cells was determined.

Boras M., Volmering S., Bokemeyer A., Rossaint J., Block H., Bardel B., Van Marck V., Heitplatz B., Kliche S., Reinhold A., Lowell C, & Zarbock A. (2017). Skap2 is required for β2 integrin–mediated neutrophil recruitment and functions. The Journal of Experimental Medicine, 214(3), 851-874.

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Macrophage Activation in Sepsis

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We prospectively enrolled 43 patients after abdominal surgery in the Intensive Care Unit of the Second Affiliated Hospital of Zhejiang University School of Medicine from August 2012 to September 2013. 27 patients received treatment for sepsis and 16 patients were recruited as nonseptic controls. All patients presented clinical and/or laboratory variables that fulfilled the criteria for sepsis [20 (link)]. The study was approved by the Human Ethics Committee of Zhejiang University and written informed consent was obtained from the patients. Macrophages were collected from the peritoneal lavage or abdominal cavity flushing fluid. The recovered cells were washed with PBS three times. Macrophages were finally resuspended in Dulbeco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA), 100 U/mL penicillin, and 100 μg/mL streptomycin (Sigma, St. Louis, MO, USA) and allowed to adhere for 2 h at 37°C, 5% CO₂. Nonadherent cells were removed by washing and the cultured cells contained >92% macrophages. Macrophages were treated with mouse anti-human MR IgG1 (5 μg/mL) or TsES (5 μg/mL) or their combination. Mouse IgG1 was the isotype control (Santa Cruz, CA, USA).

Du L., Liu L., Yu Y., Shan H, & Li L. (2014). Trichinella spiralis Excretory-Secretory Products Protect against Polymicrobial Sepsis by Suppressing MyD88 via Mannose Receptor. BioMed Research International, 2014, 898646.

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Blocking CD163 to Modulate Macrophage Phenotype

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We used a monoclonal anti-CD163 antibody (Clone RM3/1) to block functional properties of CD163, as it has been shown elsewhere in vitro in monocytes (Philippidis, et al., 2004 (link), Moura, et al., 2012 (link)), and we have successfully used to block the transition of macrophages to an anti-inflammatory-like phenotype (Alvarado-Vazquez, et al., 2017 ). In our current study we incubated skin cells co-cultured with THP-1 cells with either anti-CD163 antibody clone RM3/1 or its isotype antibody mouse IgG1 (20 μg/mL, Santa Cruz Biotechnology, Santa Cruz, CA, sc-33715, sc-3877 respectively). We applied antibody treatments two times, at 0 and 9 hours after the scratch procedure.

Ferreira D.W., Ulecia-Morón C., Alvarado-Vázquez P.A., Cunnane K., Moracho-Vilriales C., Grosick R.L., Cunha T.M, & Romero-Sandoval E.A. (2019). CD163 overexpression using a macrophage-directed gene therapy approach improves wound healing in ex vivo and in vivo human skin models.. Immunobiology, 225(1), 151862.

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Multiparametric Phenotypic Analysis of Cells

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After a 5 min dissociation step with TrypLE™Select at 37 °C, cells were washed with DPBS by centrifugation and a total of 2–3 × 10⁵ cells were used per analysis. For membrane markers, cells were incubated with the primary antibody for 1 h and with the secondary antibody for 30 min at 4 °C in DPBS with 5% (v/v) FBS. For intracellular markers, cells were permeabilized using the Inside Stain Kit (Miltenyi Biotec) according to the manufacturer‘s instructions. The following primary antibodies were used: SirPα/β (1:20, CD172a/b-PE, BioLegend), Troponin T (1:200, TnT, Thermo Scientific), CD105 (1:20,BD Pharmingen), CD166 (1:20,BD Pharmingen), CD44 (1:5, eBiosciences), CD11b (1:10, AbDSerotec), CD34 (1:5, BD Pharmingen), CD45 (1:5, BD Pharmingen), and isotype controls mouse IgG1k (1:5, BD Pharmingen), mouse IgG1 (1:2.5,Santa Cruz Biotechnologies), and rat IgG2b (1:5, eBiosciences). Cells were analyzed in a CyFlow® space (Partec GmbH) instrument, registering at least 10,000 events/sample.

Sebastião M.J., Serra M., Pereira R., Palacios I., Gomes-Alves P, & Alves P.M. (2019). Human cardiac progenitor cell activation and regeneration mechanisms: exploring a novel myocardial ischemia/reperfusion in vitro model. Stem Cell Research & Therapy, 10, 77.

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Immunohistochemical Analysis of Cartilage Markers

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After 28 days in culture (unless otherwise stated), the cells were washed once in PBS and fixed with 10% formaldehyde at 37 °C for 15 min. When fixed, the samples were permeabilised using a buffer solution (10.3 g sucrose, 0.292 g NaCl, 0.06 g MgCl₂, 0.476 g Hepes buffer, 0.5 ml Triton X, in 100 ml water, pH 7.2) at 4 °C for 5 min. The samples were then incubated at 37 °C for 5 min in 1% BSA/PBS, followed by the addition of the primary antibody (1:50 in 1% BSA/PBS, monoclonal anti-human collagen type II and aggrecan raised in mouse (IgG1), Santa Cruz Biotechnology Inc) for 1 h (37 °C). The samples were then washed in 0.5% Tween 20/PBS (5 min, ×3). A secondary, biotin-conjugated antibody (1:50 in 1% BSA/PBS, monoclonal anti-mouse (IgG), Vector Laboratories, Peterborough, UK) was added for 1 h (37 °C) followed by washing. A FITC conjugated streptavidin third layer was added (1:50 in 1% BSA/ PBS, Vector Laboratories, Peterborough, UK) at 4 °C for 30 min, and given a final wash.

Alakpa E.V., Jayawarna V., Burgess K.E., West C.C., Péault B., Ulijn R.V, & Dalby M.J. (2017). Improving cartilage phenotype from differentiated pericytes in tunable peptide hydrogels. Scientific Reports, 7, 6895.

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Renal Tissue Analysis in SLE

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Renal tissues from SLE patients and humanized SLE mice were fixed in 10% buffered formalin or cryopreserved in optimal cutting temperature (OCT) compound. Paraffin sections were prepared and stained with H&E and PAS. For immunofluorescence, frozen tissue sections were prepared and stained with rabbit anti–IL-17A (Abcam) or mouse anti–IL-17F (mouse IgG1; Santa Cruz Biotechnology) antibodies, or isotype control antibodies (that is, rabbit IgG and mouse IgG1 for IL-17A and IL-17F, respectively), followed by staining with Alexa Fluor 594–conjugated anti-rabbit IgG or Alexa Fluor 488–conjugated anti-mouse IgG, respectively. Human IgG deposition in tissues was determined on frozen sections by staining with Alexa Fluor 488–conjugated anti-human IgG (Dako). Immunofluorescence staining with isotype control antibodies is presented in the Supplementary Materials (fig. S8).

Wu H., Zhen Y., Ma Z., Li H., Yu J., Xu Z.G., Wang X.Y., Yi H, & Yang Y.G. (2016). Arginase-1–dependent promotion of TH17 differentiation and disease progression by MDSCs in systemic lupus erythematosus. Science translational medicine, 8(331), 331ra40.

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Investigating SMAD4 and MyD88 Interactions

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MyD88-deficient HEK293-I3A and Huh7 cells were transiently transfected using Lipofectamine^TM 2000 (Invitrogen, Burlington, ON, Canada) as recommended by the manufacturer with indicated plasmids. The total amount of DNA was kept constant. For assessment of SMAD4 and MyD88 interactions, MyD88 KO HEK293-I3A cells were used. Twenty-four hours after transfection, cells were lysed in 1 mL RIPA buffer containing 50 mM Tris (pH 8), 150 mM NaCl, 1% NP40, 0.5% deoxycholic acid, 0.1% SDS, and protease inhibitor cocktail (Complete Mini, Roche, Mannheim, Germany). Cell lysates were then incubated with indicated antibodies (anti-Flag, anti-HA, or anti-MyD88 antibody) for 3 h at 4°C, after which EZview Red Protein A Affinity Gel (Sigma-Aldrich) was added for another 2 h. For control reactions, mouse IgG1 (Santa Cruz) was used. The immune complexes were precipitated and washed thoroughly with RIPA buffer. Immunoprecipitated proteins were then eluted by adding sample buffer and were subsequently fractionated by SDS-polyacrylamide gel electrophoresis (PAGE) and visualized by immunoblotting with anti-Flag and anti-HA antibodies. Lysates were also immunoblotted for expression of transfected SMAD4-Flag (pRK-DPC4-Flag) and HA-MyD88 (pCMV-HA-MyD88).

Samba-Mondonga M., Calvé A., Mallette F.A, & Santos M.M. (2018). MyD88 Regulates the Expression of SMAD4 and the Iron Regulatory Hormone Hepcidin. Frontiers in Cell and Developmental Biology, 6, 105.

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