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Goat anti mouse igg fitc

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Goat anti-mouse IgG-FITC is a secondary antibody conjugated with the fluorescent label FITC (Fluorescein Isothiocyanate). It is designed to bind to mouse immunoglobulin G (IgG) antibodies, allowing for the detection and visualization of mouse-derived biomolecules in various immunological and biochemical applications.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Southern Biotech (1 mentions) Sp5 confocal microscope, by Leica (1 mentions) Multistix 10 sg, by Bayer (1 mentions) Fitc conjugated goat anti rabbit igg, by Southern Biotech (1 mentions) Digital fluorescent microscope, by Zeiss (1 mentions) Glomax explorer microplate reader, by Promega (1 mentions) Crithidia luciliae slides, by Bio-Rad (1 mentions) Inverted fluorescence microscope, by Olympus (1 mentions) Kallestad hep 2 slides, by Bio-Rad (1 mentions) Rabbit anti perilipin monoclonal antibody, by Cell Signaling Technology (1 mentions) Bx51 microscope, by Olympus (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions) Cellsence software, by Olympus (1 mentions) Hep 2 slides, by Bio-Rad (1 mentions) Dapi fluoromount g, by Southern Biotech (1 mentions)

Spelling variants of this product

Anti–mouse IgG FITC (4 citations) FITC conjugated goat anti-mouse IgG (3 citations) FITC-labeled goat anti-mouse IgG (3 citations) Goat Anti-Mouse IgG(H+L)-FITC (2 citations)

10 protocols using goat anti mouse igg fitc

1

Immunofluorescence Assay of P. falciparum

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Immunofluorescence assays (IFA) were performed as previously described [35 (link)] using salivary gland P. falciparum sporozoites. Spots were incubated with DAPI (1:50, Southern Biotech, Birmingham, AL, USA) in addition to goat anti-mouse IgG-FITC (1:100, Southern Biotech, Birmingham, AL, USA).
Schneider C.G., Taylor J.A., Sibilo M.Q., Miura K., Mallory K.L., Mann C., Karch C., Beck Z., Matyas G.R., Long C.A., Bergmann-Leitner E., Burkhard P, & Angov E. (2021). Orientation of Antigen Display on Self-Assembling Protein Nanoparticles Influences Immunogenicity. Vaccines, 9(2), 103.
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2

HEp-2 Immunofluorescence Assay Protocol

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HEp-2 immunofluorescence assays were performed as previously described [22 (link)]. In brief, serum was diluted 1/200 and then applied to HEp-2 slides (Antibodies Incorporated or BD). Staining was detected using goat anti-mouse IgG FITC at 1:500 (Southern Biotech) ANAs were scored on intensity of nuclear and cytoplasmic staining on a scale of 0–3 and for the presence or absence of mitotic chromatin using wide-field fluorescence microscopy (Olympus IX83; Cellsense software). The scorer used blinded to the genotype and gender of the mice.
Gordon R.A., Tilstra J.S., Marinov A., Nickerson K.M., Bastacky S.I, & Shlomchik M.J. (2020). Murine lupus is neutrophil elastase-independent in the MRL.Faslpr model. PLoS ONE, 15(4), e0226396.
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3

IFA Profiling of CP4 Bacteria Antibody Binding

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For IFA experiments, CP4 bacteria were collected from 1 ml overnight TPG culture by centrifugation at 16,000 × g for 10 min and washed twice with PBS. The pellets were resuspended with 1 ml PBS and incubated with pooled sera from MV immunized mice at the dilution of 1:400, whereas sera from BSG control mice were used as the negative control. After incubation at room temperature for 1 h, the bacteria were washed twice with PBS and then incubated with PBS containing goat anti-mouse IgG-FITC (Southern Biotech) at the dilution of 1:1000 at room temperature for 1 h. After additional twice PBS washes, the pellets were resuspended using 100 µl PBS and 10 µl samples were used for microscope analysis using a Leica SP5 confocal microscope (Germany) with a 100 × oil immersion optic.
Jiang Y., Kong Q., Roland K.L, & Curtiss R I.I.I. (2014). Membrane vesicles of Clostridium perfringens Type A strains induce innate and adaptive immunity. International journal of medical microbiology : IJMM, 304(0), 431-443.
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4

Kidney histology and immunofluorescence analysis

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Urine samples were collected weekly or before euthanasia and analyzed as described before (19 (link)) for proteinuria using Multistix 10 SG (Bayer). Kidney sections fixed in 10% buffered formalin were paraffin embedded and stained with Hematoxylin and Eosin (H & E) at the University of Virginia Research Histology Core. The sections were scored in a blinded manner on a scale of 0 to 4 for abnormalities (glomerular hypertrophy, mesangial expansion, mononuclear infiltrates and tubulo-interstitial inflammation), as 0 (no abnormality), 1 (less than 20%), 2 (20 to 40%), 3 (40 to 60%), and 4 (more than 60%) of 200x magnified field with above-mentioned abnormalities. IgG, C3, IgG2a and IgG2b deposition in kidneys was analyzed by direct immunofluorescence as before (19 (link)). Briefly, frozen sections of mouse kidneys were fixed in methanol at −20°C for 10 min, blocked with 2% normal goat serum in PBS containing 3% BSA, and then stained with goat anti mouse IgG-FITC, goat anti mouse IgG2a-TXRD, goat anti mouse IgG2b-TXRD (Southern Biotech) and goat anti-mouse Complement C3-FITC (MP Biomedicals). The frozen sections stained for immune complex deposits were analyzed by immunofluorescence captured in 200x amplified images from at least 30 random glomeruli for each group and quantitated using ImageJ software and expressed as total immunofluorescence.
Stremska M.E., Dai C., Venkatadri R., Wang H., Sabapathy V., Kumar G., Jose S., Mohammad S., Sung S.S., Fu S.M, & Sharma R. (2019). IL233, an IL-2-IL-33 Hybrid Cytokine Induces Prolonged Remission of Mouse Lupus Nephritis by Targeting Treg cells as a Single Therapeutic Agent. Journal of autoimmunity, 102, 133-141.
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5

Immunohistochemical Analysis of Renal IgG Deposition and SOCS1 Expression

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As described previously (8 (link)), IHC was performed for renal IgG deposition. For detection of SOCS1, paraffin sections were incubated with rabbit anti-mouse SOCS1 (2 µg/ml; Abcam, Cambridge, MA, USA), and then with polymer-horseradish peroxidase conjugated goat anti-rabbit IgG (1 µg/ml; DAKO, Glostrup, Denmark). A brown color was developed with 3,30-diaminobenzidine chromogen substrate (DAKO).
Immunofluorescence was carried out for both IgG deposition and SOCS1 expression in frozen sections. Routinely processed sections were incubated with rabbit anti-mouse SOCS1 and then fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG (2 µg/ml; Southern Biotech, Birmingham, AL, USA) in order, or directly with goat anti-mouse IgG-FITC (Southern Biotech) alone. Sections were viewed under a digital fluorescent microscope (Carl Zeiss, Jena, Germany).
Wang P., Yang J., Tong F., Duan Z., Liu X., Xia L., Li K, & Xia Y. (2017). Anti-Double-Stranded DNA IgG Participates in Renal Fibrosis through Suppressing the Suppressor of Cytokine Signaling 1 Signals. Frontiers in Immunology, 8, 610.
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6

Histological Assessment of Glomerulonephritis

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Four weeks after the first HgCl2 injection, kidneys were harvested from mice and embedded in Tissue-Tek OCT compound (Sakura, Torrance, CA) and snap-frozen on dry ice. A cryostat microtome was used to cut 4 µm sections. The sections were air dried, fixed with cold acetone and stained with goat anti-mouse IgG FITC (Southern Biotechnology Associates, Birmingham, AL) to detect IgG deposits. Glomerulonephritis was scored in a blinded fashion using 1–4 scale based on the percentage of glomeruli affected. The grading scale was 0 =  no glomerulonephritis, 1+ ≤15%, 2+16–40%, 3+41–70%, and 4+ >70% of glomeruli affected. The lesions graded included thickening of the mesangium, noticeable increases in both mesangial and glomerular cellularity with/without superimposed inflammatory exudates and capsular adhesions, glomerulosclerosis, and cast formation. Scoring was performed on at least 200 glomeruli in a 40× field per kidney [32] (link). Immune complex deposition in kidneys was quantified by blinded observers and arbitrarily scored from 0 to 4 dependent on the intensity and amount of the anti-IgG staining in the glomeruli [32] (link).
Jha V., Workman C.J., McGaha T.L., Li L., Vas J., Vignali D.A, & Monestier M. (2014). Lymphocyte Activation Gene-3 (LAG-3) Negatively Regulates Environmentally-Induced Autoimmunity. PLoS ONE, 9(8), e104484.
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7

Quantification of Anti-dsDNA Autoantibodies

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Serum and fecal anti-dsDNA IgG, IgM and IgA were quantified by ELISA with 1:100 dilutions as previously described14 (link). Anti-dsDNA IgG were also visualized on Crithidia luciliae slides (Bio-Rad) with serum diluted 1:10 followed by an incubation with goat anti-mouse IgG-FITC (1:100; Southern Biotech). Images were acquired on an inverted fluorescence microscope (Olympus). The corrected total cell fluorescence (https://theolb.readthedocs.io/en/latest/#) from each Crithidia luciliae cell was quantified with ImageJ. IL-6 or IFN-γ were quantitated with BD Biosciences ELISA kits in serum samples diluted 1:50 assayed in duplicate. The absorbance was detected with the Promega GloMax® Explorer microplate reader at 450 nm.
Ma L., Ge Y., Brown J., Choi S.C., Elshikha A., Kanda N., Terrell M., Six N., Garcia A., Mohamadzadeh M., Silverman G, & Morel L. (2024). Dietary tryptophan and genetic susceptibility expand gut microbiota that promote systemic autoimmune activation. bioRxiv.
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8

Autoantibody Profiling in Murine Models

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Proteinuria, glomerulonephritis, interstitial renal disease, and dermatitis were evaluated exactly as previously described [11 ] by individuals blinded to the genotypes of the animals.
HEp-2 antinuclear antibody assays were performed using Kallestad HEp-2 slides (Bio-Rad, Hercules, CA) as a substrate. Serum samples were diluted to 1/100 in 1x PBS, 1% BSA, 0.05% sodium azide, and bound autoantibodies were detected using goat anti-mouse IgG-FITC (Southern Biotech, Birmingham, AL). Samples were scored for the dominant staining pattern, presence or absence of mitotic chromatin staining, and relative cytoplasmic staining intensity by an individual blinded to the genotypes of the samples.
Autoantibody and total IgM and IgG ELISAs were performed essentially as previously described [11 ]. Samples were assayed over a total of eight three-fold dilutions starting at 1/100 for anti-nucleosome and anti-Sm ELISAs, four three-fold dilutions starting at 1/100 for anti-RNA and anti-IgG2a rheumatoid factor ELISAs, and eight three-fold dilutions starting at 1/5000 for total IgM ELISA or 1/50000 for total IgG ELISA. ELISA standards were purified PL2-3 and BWR4 antibodies, hybridoma supernatants from Y2 and 400tμ23 cultures, or purified unconjugated IgM,κ and IgG1,κ isotype control antibodies (BD Biosciences, Franklin Lakes, NJ).
Nickerson K.M., Wang Y., Bastacky S, & Shlomchik M.J. (2017). Toll-like receptor 9 suppresses lupus disease in Fas-sufficient MRL Mice. PLoS ONE, 12(3), e0173471.
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9

Adipocyte Immunofluorescence Staining Protocol

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Optimal cutting temperature (OCT) medium embedded SCAT and OMAT were cut at 5- to 7-μm thickness and mounted on gelatin-coated slides as previously described [23 (link)]. The sections were fixed with methanol and acetone mixture (1:1), and then the first and second primary antibodies, rabbit anti-perilipin monoclonal antibody (the marker for adipocyte, Cell signaling, Beverly, MA), goat polyclonal ADM and CRLR antibody (Santa Cruz Biotechnology Inc., Dallas, TX), and rabbit RAMP2 and RAMP3 polyclonal antibodies [24 (link)] were applied at 1:1000 dilution, followed by either goat anti-mouse IgG-FITC (Southern Biotech, Birmingham, AL), or donkey anti-goat Alexa Flour 594/anti-rabbit IgG TRITC (Life Technology, Grand Island, NY). The slides were then mounted with mounting-medium containing 4′, 6-diamidino-2-phenylindole (DAPI; Vector Laboratories Inc., Burlingame, CA) and viewed under an Olympus BX51 microscope. The intensity of the immunofluorescence was measured by using CellSence software (Olympus Scientific, Walthan MA, USA), and the relative densities of the immunofluorescence to the number of nuclei were calculated and compared between groups.
Dong Y., Betancourt A., Belfort M.A, & Yallampalli C. (2022). Lipid dysfunction and adrenomedullin expression in omental versus subcutaneous adipose tissues in diabetic pregnancies. PLoS ONE, 17(4), e0265419.
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10

Detecting Autoantibodies in Murine Plasma

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Autoantibodies were assessed using Hep‐2 slides (BioRad). Briefly, slides were incubated with plasma of PBS or pristane‐treated mice diluted 1:50 in PBS in a humidified chamber at room temperature for 30 min. The slides were quickly rinsed in PBS followed by two five minute washes in PBS. Goat anti‐mouse IgG FITC (Southern Biotech) at a 1:100 dilution in PBS was then added to the slides. The slides were incubated in a humidified chamber at room temperature for 30 min, protected from light. After washing two times with PBS, several drops of DAPI Fluoromount G (Southern Biotech) were added to the slides, followed by a coverslip. The slides were visualized using a Lionheart Pfx microscope (BioTek).
McClung D.M., Kalusche W.J., Jones K.E., Ryan M.J, & Taylor E.B. (2021). Hypertension and endothelial dysfunction in the pristane model of systemic lupus erythematosus. Physiological Reports, 9(3), e14734.
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