Mouse igg1 isotype control

Manufactured by R&D Systems

Sourced in United States, United Kingdom, Japan

The Mouse IgG1 isotype control is a laboratory reagent used as a negative control in flow cytometry, immunohistochemistry, and other immunoassays. It is a purified mouse immunoglobulin G1 (IgG1) antibody with no known reactivity to human antigens.

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Lab products found in correlation

Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Facsdiva software, by BD (2 mentions) Attune nxt software v2, by Thermo Fisher Scientific (2 mentions) Attune next flow cytometer, by Thermo Fisher Scientific (2 mentions) Sp600125, by Merck Group (2 mentions) Mab002, by R&D Systems (2 mentions) Rpmi 1640, by Thermo Fisher Scientific (2 mentions) Rneasy mini kit, by Qiagen (1 mentions) Hprt1 hs02800695 m1, by Thermo Fisher Scientific (1 mentions) Aldefluor kit, by STEMCELL (1 mentions) Exosome depleted fbs, by System Biosciences (1 mentions) Facscalibur, by BD (1 mentions) Alexa fluor 647, by Thermo Fisher Scientific (1 mentions) Facscanto 2 cytometer, by BD (1 mentions) Lyoplate screening panel, by BD (1 mentions) Tunicamycin, by Merck Group (1 mentions) Tunicamycin, by R&D Systems (1 mentions) Pepinh myd, by InvivoGen (1 mentions) Anti dap12, by Cell Signaling Technology (1 mentions) Isotype control, by BioXCell (1 mentions)

Spelling variants of this product

Isotype control (26 citations) Mouse IgG1 (22 citations) IgG1 isotype control (15 citations) Mouse IgG1 isotype control antibody (3 citations) Isotype control mouse antibodies for IgG1 and IgM (2 citations) Mouse IgG1 Isotype control (11711) (2 citations) Mouse monoclonal IgG1 isotype control (2 citations) Phycoerythrin-conjugated mouse IgG1 isotype control (2 citations) Mouse IgG1 PE isotype control (1 citations)

32 protocols using mouse igg1 isotype control

Quantification of PAD-2 and PAD-4 Expression

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RA PBMCs and SFMCs were stimulated with either 10 ng/ml TNFα (Peprotec), 800 ng/ml rhPD-L2/B7-DC Fc chimera protein (R&D Systems), or 800 ng/ml Mouse IgG1 Isotype control (R&D Systems) for 2 h at 37 °C and 5% CO_2, at a cell density of 1–2.5 × 10⁶ cells/ml.
RNA was extracted using the RNeasy mini kit (Qiagen) according to manufacturer’s instructions. The concentration of RNA was determined using the Nanodrop 1000 Spectrophotometer. Followed by real-time PCR analysis using the Taqman RNA to Ct 1-step kit (Applied Bioscience) and the following TaqMan assays: PAD-2 (Hs00247108_m1), PAD-4 (Hs00202612_m1), and HPRT1 (Hs02800695_m1) (all from ThermoFisher Scientific). The relative amount of PAD-2 and PAD-4 mRNA were calculated using the formula: 2^-ΔCq with (Hypoxanthine-guanine-phosphoribosyl-transferase 1) HPRT1 as the reference gene, and normalized to the N/A sample (not-activated) to obtain a relative ratio.

Greisen S.R., Kragstrup T.W., Thomsen J.S., Hansen A.S., Krishnamurthy A., Hørslev-Petersen K., Hetland M.L., Stengaard-Pedersen K., Østergaard M., Ørnbjerg L.M., Junker P., Sharpe A.H., Freeman G.J., Annamalai L., Hvid M., Moestrup S.K., Hauge E.M., Catrina A.I, & Deleuran B. (2019). Programmed death ligand 2 – A link between inflammation and bone loss in rheumatoid arthritis. Journal of Translational Autoimmunity, 3, 100028.

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EpCAM and ALDH1 Analysis in Sorted Cells

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EpCAM^high cell percentages within the AC133 – and + sorted-cells, were analyzed by flow cytometry (FCM) from 5.10⁴ cells. After saturation in 1% BSA in DPBS 1X calcium and magnesium free (10min, 4°C), cells were incubated for 45min at room temperature (RT) with an EpCAM mouse monoclonal Ab (clone VU1D9; Ozyme - Cell Signaling Technology, France) diluted at 1:150 in 1% BSA/DPBS. After a washing step in DPBS 1X (g x 300, 10min, 4°C), cells were incubated for 30min at RT in the dark with an Alexa-Fluor 633-conjugated goat anti-mouse secondary Ab (ThermoFisher Scientific, France) diluted at 1:1000 in DPBS 1X.
Enzymatic activity of ALDH1 in MIX+ and MIX- sorted-cells was analyzed from 10⁵ cell/mL, using the ALDEFLUOR kit (Stem Cell Technologies, France) according to the manufacturer’s recommendations.
Cells were extemporaneously stained with a DNA dye, i.e., 0.5µL propidium iodide (PI; λex=475-581nm/λem=583-697nm; BD Biosciences, France). EpCAM^high immunostaining and ALDH1 enzymatic activity (ALDH1bright cells) were analyzed among live cells (PI-), with the BD AccuriC6 Plus FCM (BD Biosciences, France). Mouse IgG1 isotype control (R&D systems, France) and N, N-diethylaminobenzaldehyde (DEAB, ALDH1 specific inhibitor) were used to control for background fluorescence

Blondy S., Durand S., Lacroix A., Christou N., Bouchaud C., Peyny M., Battu S., Chauvanel A., Carré V., Jauberteau M.O., Lalloué F, & Mathonnet M. (2022). Detection of Glycosylated Markers From Cancer Stem Cells With ColoSTEM Dx Kit for Earlier Prediction of Colon Cancer Aggressiveness. Frontiers in Oncology, 12, 918702.

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CM Stimulation and sEV Isolation

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The CM was collected and used for the stimulation of other cells or for Western blotting. CM was centrifuged at 1500 rpm for 3 min to remove cell debris, and the supernatant was used in the subsequent experiments. For Western blotting, MDA-MB-231 and MDA-MB-453 cells were cultured in a serum-free RPMI-1640 medium 24 h before CM collection so as not to carry over the proteins from the FBS. For sEV isolation, these cells were cultured using exosome-depleted FBS (System Biosciences, Palo Alto, CA, USA). For the neutralization of HSP70, the CM was preincubated with the HSP70 antibody (Table S1) or a mouse IgG1 isotype control (5 μg/mL, R&D Systems Inc., Minneapolis, MN, USA) for 1 h at 37 °C, and the THP-1-derived macrophages and MDA-MB-231 cells were then treated with the CM.

Yamaguchi-Tanaka M., Takagi K., Miki Y., Sato A., Iwabuchi E., Miyashita M, & Suzuki T. (2023). The Pro-Tumorigenic Role of Chemotherapy-Induced Extracellular HSP70 from Breast Cancer Cells via Intratumoral Macrophages. Cancers, 15(6), 1903.

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Flow Cytometric Analysis of CD302 Expression

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Cells were trypsinized, pelleted, and resuspended in FACS buffer (1% FCS in PBS). Cells were treated with PBS containing 1% FCS and 0.5% saponin for 20 min on ice for permeabilization. Unconjugated monoclonal antibody specific for CD302 (human CD302/CLEC13A; R&D Systems; monoclonal mouse immunoglobulin G1 [IgG1]) or a mouse IgG1 isotype control (R&D Systems, Minneapolis, MN, USA) were diluted in FACS buffer or permeabilization buffer to 5 μg/mL and added to cells for 30 min at 4°C. Following two washes with FACS buffer, a specific anti-mouse secondary antibody conjugated to Alexa Fluor 488 or Alexa Fluor 647 (Thermo Fisher Scientific) was diluted 1:1,000 in FACS buffer or permeabilization buffer and added to cells. After three washes with FACS buffer, stained cells were resuspended in FACS buffer or fixation buffer (1% FCS and 0.5% paraformaldehyde [PFA] in PBS) and analyzed by BD FACSCalibur (BD Biosciences, Franklin Lakes, NJ, USA) or SONY SA3800 spectral analyzer. Data were analyzed using FlowJo software (BD Biosciences).

Reinecke B., Frericks N., Lauber C., Dinkelborg K., Matthaei A., Vondran F.W., Behrendt P., Haid S., Brown R.J, & Pietschmann T. (2022). The Human Liver-Expressed Lectin CD302 Restricts Hepatitis C Virus Infection. Journal of Virology, 96(7), e01995-21.

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Evaluation of MET and CXCR4 Receptors in RMS

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For evaluation of MET and CXCR4 receptors expression levels RMS cells were stained with monoclonal FITC-labeled anti-human HGFR/c-MET antibody, clone 95106 (R&D), PE-labeled anti-human CXCR4 antibody (Becton Dickinson, Franklin Lakes, NJ, USA) or mouse IgG1 isotype control (R&D) labeled with FITC or PE respectively. The cells were acquired using FACS Canto II cytometer (Becton Dickinson) and analyzed using FACS Diva software (Becton Dickinson), as described previously [21 (link)].
The expression level of adhesion molecules was evaluated using Lyoplate technology (Lyoplate Screening Panel, Becton Dickinson) according to the manufacturer’s protocol. The cells were acquired by use of Attune Next Flow Cytometer and analyzed using Attune NxT Software v2.2 (Thermo Fisher Scientific, Waltham, MA, USA).

Skrzypek K., Kot M., Konieczny P., Nieszporek A., Kusienicka A., Lasota M., Bobela W., Jankowska U., Kędracka-Krok S, & Majka M. (2020). SNAIL Promotes Metastatic Behavior of Rhabdomyosarcoma by Increasing EZRIN and AKT Expression and Regulating MicroRNA Networks. Cancers, 12(7), 1870.

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Capturing SARS-CoV-2 Virions with Antibodies

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The following mouse and human monoclonal antibodies were utilized for virion capture assays: anti-PSGL-1 and mouse IgG1 isotype control (BD Biosciences, Cat#556053 and 557273) and anti-gp120s (clones 2G12 and PG9 acquired from the NIH ARP; Cat#1476 and 12149). For virion capture assays in patient plasmas, we used biotinylated antibodies, including CD44-bio (Miltenyi, Cat#130-113-340) and an in-house biotin conjugation (Bio-Rad Laboratories, Cat#LNK262B) of anti-PSGL-1 and mouse IgG1 isotype control (R&D, Cat#MAB002) antibodies. PE-conjugated mouse anti-PSGL-1 mAb (BD Biosciences, Cat#556055) was used for flow virometry labelling.

Burnie J., Persaud A.T., Thaya L., Liu Q., Miao H., Grabinsky S., Norouzi V., Lusso P., Tang V.A, & Guzzo C. (2022). P-selectin glycoprotein ligand-1 (PSGL-1/CD162) is incorporated into clinical HIV-1 isolates and can mediate virus capture and subsequent transfer to permissive cells. Retrovirology, 19, 9.

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Tunicamycin and CA 19.9 MAb Effects on NCI-N87 Cells

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Following trypsinization of confluent NCI-N87 cultures, 5 × 10⁵ cells were seeded in 6-well plates and allowed to attach for 24 h. Cells were then incubated for another 24 h with 0.01, 0.1 and 1 µg/mL of tunicamycin (Sigma-Aldrich) and 0.2 and 2 µg/mL of CA 19.9 MAb (Santa Cruz Biotechnology), in RPMI 1640 GlutaMAX supplemented with 10% heat-inactivated fetal bovine serum and simple RPMI 1640 GlutaMAX, respectively. DMSO (0.05%) and 2 µg/mL of mouse IgG1 isotype control (R&D Systems) were used as controls for tunicamycin and CA 19.9 treatment, respectively. Regarding CA 19.9 treatment, an additional control of complete growth medium was included. Finally, total protein extracts were collected for each experimental condition and WB was performed as described in Section 4.2. All experiments were performed in duplicate.

Duarte H.O., Balmaña M., Mereiter S., Osório H., Gomes J, & Reis C.A. (2017). Gastric Cancer Cell Glycosylation as a Modulator of the ErbB2 Oncogenic Receptor. International Journal of Molecular Sciences, 18(11), 2262.

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Monoclonal Antibodies for Human LSECtin

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mAbs to human LSECtin were established by immunization of Balb/C mice with recombinant LSECtin extracellular domain protein. Three independent clones, CCA023 (IgG2a), CFD051 (IgG1) and CCB059 (IgG2b), were established [34 (link)]. The anti-human LSECtin mAb CCB059 (IgG2b) was selected for staining by flow cytometry. The mouse IgG1 isotype control was from R&D Systems (Minneapolis, MN, USA). mAbs against human HLA-DR, CD83 and CD86 were from eBioscience (San Diego, CA, USA); mAbs against human CD40 and CD80 were from Biolegend (San Diego, CA, USA); anti-phosphotyrosine Ab (4G10) was from Millipore; anti-DAP12 and the other phospho-specific Abs were from Cell Signaling Technology (Danvers, MA). The Syk inhibitors piceatannol and R406 were purchased from Calbiochem (San Diego, CA, USA) and Selleckchem (Houston, TX, USA) respectively. Raf-1 inhibitor GW5074 was purchased from Calbiochem (San Diego, CA, USA); The MyD88 inhibitory peptide Pepinh-MYD was from InvivoGen (San Diego, CA, USA). The GlcNAc β1-2Man disaccharide was purchased from Dextra Laboratories (Reading, UK).

Zhao D., Han X., Zheng X., Wang H., Yang Z., Liu D., Han K., Liu J., Wang X., Yang W., Dong Q., Yang S., Xia X., Tang L, & He F. (2016). The Myeloid LSECtin Is a DAP12-Coupled Receptor That Is Crucial for Inflammatory Response Induced by Ebola Virus Glycoprotein. PLoS Pathogens, 12(3), e1005487.

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Modulating Inflammation in Colorectal Cancer

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For SP600125 treatment, WT and Rip3^-/- mice were administered the JNK inhibitor SP600125 (40 mg/kg) (Sigma-Aldrich) or vehicle (DMSO) by oral gavage once daily during CAC induction. For anti-CXCL1 treatment, WT and Rip3^-/- mice receiving AOM/DSS were treated either with a mouse anti-CXCL1 neutralizing antibody (120 µg/mouse once a week, i.p.) or mouse IgG1 isotype control (both from R&D Systems) until sacrifice (day 100). For anti-CD90 treatment, T cells were depleted with neutralizing anti-CD90 monoclonal antibody as previously described 23 (link). Rip3^-/- mice receiving AOM/DSS were treated either with a mouse anti-CD90 neutralizing antibody (100 µg/mouse once a week, i.p.) or isotype control (both from BioXcell) until sacrifice (day 100). For recombinant CXCL1 treatment, Rip3^-/- mice receiving AOM/DSS were treated either with mouse recombinant CXCL1 (300 ng/mouse twice a week, i.p.) or control PBS (both from R&D Systems) until sacrifice (day 100).

Liu Z.Y., Zheng M., Li Y.M., Fan X.Y., Wang J.C., Li Z.C., Yang H.J., Yu J.M., Cui J., Jiang J.L., Tang J, & Chen Z.N. (2019). RIP3 promotes colitis-associated colorectal cancer by controlling tumor cell proliferation and CXCL1-induced immune suppression. Theranostics, 9(12), 3659-3673.

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TACSTD2 Gene Overexpression and Silencing

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A pcDNA3.1 vector carrying the TACSTD2 gene insert and a pcDNA3.1 vector carrying the siRNA-resistant TACSTD2 gene devoid of the 3’- and 5’-untraslated regions (UTR) (pTACSTD2) for rescue experiments were purchased from GenScript Corporation. siRNA for TACSTD2 gene silencing (siTACSTD2) targeting the 3’UTR of the TACSTD2 gene was performed using SMARTpool of 4 siRNAs (Dharmacon) (siTD2-A, 5’-GAGAAGAGGAGUUUGUUAAUU-3’; siTD2-B, 5’-ACAAGUAUCUGUAUGACAAUU-3’, siTD2-C, 5’-GCAAGUAACUGAAUCCAUUUU-3’, siTD2-D, 5’-GCACACACCAGGUUUAAUAUU-3’), which were transfected into parental and TACSTD2-overexpressing Huh 7.5 cells at 100nM final concentration using RNAiMAX (Invitrogen). The antibodies used include mouse anti-TACSTD2 (Santa Cruz), goat anti-TACSTD2, biotinylated goat anti-TACSTD2 and goat anti-E-cadherin, mouse IgG1 isotype control, mouse IgG2A isotype control, and normal goat IgG control (R&D systems), mouse anti-CLDN1 (Abnova), rabbit anti-CLDN1 and mouse anti-OCLN (Life Technologies), rabbit anti-OCLN (Abcam), rabbit anti-CD81 and mouse anti-ZO-1 (Thermo Scientific), mouse anti-SR-B1 (BD transduction laboratories), mouse anti-CD81 (BD Pharmingen), mouse anti-JAM-A (Hycult Biotech), mouse anti-HCV core (Anogen), rabbit anti-PKC substrate (Cell Signaling Technologies), and rabbit anti-alpha tubulin (Millipore). The HCV neutralizing mAb AR4A was a gift from Dr. Mansun Law.

Sekhar V., Pollicino T., Diaz G., Engle R.E., Alayli F., Melis M., Kabat J., Tice A., Pomerenke A., Altan-Bonnet N., Zamboni F., Lusso P., Emerson S.U, & Farci P. (2018). Infection with hepatitis C virus depends on TACSTD2, a regulator of claudin-1 and occludin highly downregulated in hepatocellular carcinoma. PLoS Pathogens, 14(3), e1006916.

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