Anti mouse antibodies

Manufactured by Thermo Fisher Scientific

Sourced in United States

Anti-mouse antibodies are laboratory reagents used to detect and analyze mouse proteins or other biomolecules in various experimental settings. They provide a specific and sensitive tool for identifying and quantifying mouse-derived targets in biological samples.

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Lab products found in correlation

Cell strainer, by BD (1 mentions) Permeabilization solution, by Thermo Fisher Scientific (1 mentions) Brefeldin a, by Thermo Fisher Scientific (1 mentions) Ionomycin, by Merck Group (1 mentions) Facscanto 2, by BD (1 mentions) Anti il 4 pe, by Thermo Fisher Scientific (1 mentions) Annexin 5 fitc propidium iodide apoptosis kit, by Beyotime (1 mentions) Collagenase type 2, by Merck Group (1 mentions) Anti cd206 pe, by Thermo Fisher Scientific (1 mentions) Cd4 t cell isolation kit, by Miltenyi Biotec (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Moflo xdp cell sorter, by Beckman Coulter (1 mentions) System 2 software, by Beckman Coulter (1 mentions) Mouse t cell negative selection kit, by STEMCELL (1 mentions) Propidium iodide, by Merck Group (1 mentions) Bovine serum albumin (bsa), by Gemini Bio (1 mentions) Anti gapdh, by Thermo Fisher Scientific (1 mentions) Pvdf membrane, by Merck Group (1 mentions) 4 6 diamino 2 phenylindole dapi, by Thermo Fisher Scientific (1 mentions) Carl microscope, by Zeiss (1 mentions)

Close spelling variants of this product

Anti-mouse secondary antibodies Anti-rabbit and anti-mouse secondary antibodies Alexa Flour donkey anti‐rabbit and donkey anti‐mouse antibodies Alexa488- or Alexa594-conjugated anti-mouse or anti-rabbit antibodies Anti-rabbit or anti-mouse IgG antibodies DyLight anti-mouse and anti-rabbit antibodies Alexa 488/568-conjugated donkey anti-mouse/rabbit secondary antibodies Fluorescently tagged anti-rabbit or anti-mouse secondary antibodies Anti-mouse monoclonal antibodies HRP-linked secondary anti-mouse or anti-rabbit antibodies

10 protocols using anti mouse antibodies

Neutrophil Analysis in Colon Lamina Propria

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The lamina propria of the colon was collected and processed for neutrophil analysis. First, the colon was extracted and washed twice with 1× HBSS supplemented with 5% FBS and 2 mM EDTA in a 50-ml Falcon tube under horizontal shaking in an orbital mixer at 250 rpm for 20 min at 37°C. Then, enzymatic digestion was performed in solution containing 1× HBSS supplemented with 2 mM EDTA and collagenase IV (40 U/ml) at 200 rpm for 15 min at 37°C. The obtained cells were then separated from the tissue using a 70-µm filter (Cell Strainer; BD Bioscience). Samples were counted in Neubauer’s chamber, and 10⁶ cells were labeled with specific antibodies coupled to different fluorochromes and analyzed by flow cytometry. Anti-Ly6G FITC (Gr-1) clone 1A8-Ly6g, anti-Ly6C PE clone HK1.4, and anti-CD11b APC clone M1/70 anti-mouse antibodies (eBioscience, Thermo Fisher Scientific) were used for analysis of the neutrophil population on the lamina propria. Data were analyzed using FlowJo software.

Fachi J.L., Sécca C., Rodrigues P.B., de Mato F.C., Di Luccia B., Felipe J.D., Pral L.P., Rungue M., Rocha V.D., Sato F.T., Sampaio U., Clerici M.T., Rodrigues H.G., Câmara N.O., Consonni S.R., Vieira A.T., Oliveira S.C., Mackay C.R., Layden B.T., Bortoluci K.R., Colonna M, & Vinolo M.A. (2019). Acetate coordinates neutrophil and ILC3 responses against C. difficile through FFAR2. The Journal of Experimental Medicine, 217(3), jem.20190489.

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Splenocyte Immunophenotyping and Cytokine Profiling

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Single cell suspensions were obtained from spleens of recipient mice. Cells were labeled for surface and intracellular antigens with fluorescence-labeled anti-mouse antibodies (eBioscience, CA). Intracellular staining for Foxp3 was performed with a commercially available staining kit including permeabilization solution and buffer (eBioscience, CA). For intracellular cytokine staining, total splenocytes were seeded on 96-well plates and stimulated in complete media for 4 hrs at 37°C with phorbol 12-myristate 13-acetate (PMA; 50 ng/ml; Sigma-Aldrich), Ionomycin (500ng/ml; Sigma-Aldrich) and Brefeldin A (eBioscience). Subsequently, cells were fixed, permeabilized (BD Biosciences, CA) and stained with respective antibodies. Flow cytometry measurements were performed on a FACS Canto II (BD Bioscience, CA) and data were analyzed using FlowJo (FlowJo Software, OR).

Heinbokel T., Quante M., Iske J., Nian Y., Maenosono R., Minami K., Liu Y., Azuma H., Elkhal A, & Tullius S.G. (2020). CTLA4-Ig prolongs graft survival specifically in young but not old mice. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 21(2), 488-502.

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Cardiac Cell Isolation and Flow Cytometry

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After 7 days, cells were collected from the culture flasks by scraping and resuspended in PBS. After washed twice, cells were centrifuged at 300 g for 5 min and resuspended in 100 µl PBS. Then cells were incubated with antibodies or corresponding IgG controls for 30 min on ice.
For cardiac cell collection, the hearts were dissected, minced and enzymatically digested with type II collagenase (Sigma, USA), and passed through a 70-µm cell strainer. The following anti-mouse antibodies (eBioscience, USA) were used: anti-CCR2-APC, anti-CD206-PE, anti-CD3-FITC anti-CD4-APC, and anti-IL-4-PE. Data were analyzed with FlowJo software (Treestar, USA).
The H9c2 cardiomyocytes (Rat cardiomyocytes; American Type Culture Collection, Manassas, USA) were cultured in DMEM with 10% FBS and 1% P/S. The culture conditions contained 95% air and 5% CO₂ at 37 ℃. An Annexin V-FITC/propidium iodide (PI) apoptosis kit (Beyotime, China) was used to analyze the apoptosis rate. The early apoptosis was gated as Annexin V⁺/PI⁻ population while late apoptosis was gated as Annexin V⁺/PI⁺ population.

Liu Y., Wu M., Zhong C., Xu B, & Kang L. (2022). M2-like macrophages transplantation protects against the doxorubicin-induced heart failure via mitochondrial transfer. Biomaterials Research, 26, 14.

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Isolation of Naïve CD4+ T Cells

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Wild type or Tpl2^−/− cells from spleens and lymph nodes were disaggregated by pressing through a 70 μm filter, and CD4 T cells were column purified by negative selection using a CD4⁺ T cell isolation kit according to manufacturer’s guidelines (Miltenyi Biotech, Auburn, CA). CD4 T cells were stained for 15 min at 4°C in PBS + 0.5% FBS (Life Technologies, Carlsbad, CA) using anti-mouse antibodies purchased from eBioscience (San Diego, CA): CD16/CD32 (93), CD4 (RM4-5), TCRβ (H57-597), CD25 (PC61.5), CD44 (IM7), CD62L (MGL-14), and CD45RB (C363.16A). Live cells were first gated by excluding propidium iodide positive (PI⁺) cells and then sorted for naïve effectors (CD4⁺CD25^-CD62L⁺CD44⁻ or CD4⁺CD25⁻CD45RB^hi) using a Beckman Coulter MoFlo XDP cell sorter.

Acuff N.V., Li X., Kirkland R., Nagy T, & Watford W.T. (2015). Tumor Progression Locus 2 Differentially Regulates IFNγ and IL-17 Production by Effector CD4+ T Cells in a T Cell Transfer Model of Colitis. PLoS ONE, 10(3), e0119885.

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Flow Cytometry Analysis of T-Cell Subsets

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The percentage of CD4⁺ T cells and CD8⁺ T cells present in the spleens were determined by using flow cytometry according to our previous study [30 (link)]. The CD3, CD4, and CD8 antigens on the surfaces of the T-cell subclasses were stained with the corresponding anti-mouse antibodies (eBioscience) tagged with phycoerythrin (PE), allophycocyanin (APC), and fluorescein isothiocyanate (FITC), respectively. The fluorescence profiles of all the samples were analyzed on a FACScan flow cytometer (BD Biosciences) using the System II Software (Coulter).

Zhang N.Z., Xu Y., Wang M., Chen J., Huang S.Y., Gao Q, & Zhu X.Q. (2016). Vaccination with Toxoplasma gondii calcium-dependent protein kinase 6 and rhoptry protein 18 encapsulated in poly(lactide-co-glycolide) microspheres induces long-term protective immunity in mice. BMC Infectious Diseases, 16, 168.

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Multilineage Differentiation of MSCs

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MSCs were characterized by flow cytometry using anti-mouse antibodies (all from eBioscience) directed to cell surface markers Sca1, CD44, CD73, CD45, CD29, and CD 105, together with relevant isotype controls. Recombinant mouse E-selectin (CD62E)-human Fc chimera (“mE-Ig”) was purchased from R&D Systems. MSCs were tested for their differentiation capacity into various mesodermal lineages as previously described [21 (link), 22 (link)]. Briefly, chondrogenic differentiation of MSCs was induced with ascorbic acid (50 μg/ml) and TGFβ1 (1 ng/ml) in culture medium. After 3 weeks of culture, plates were washed with PBS, fixed with 4% paraformaldehyde, and stained with 0.05% alcian blue for light microscopic visualization of cartilage. Osteogenic differentiation was induced with ascorbic acid (50 μg/ml), sodium β-glycerophosphate (10 mM), and dexamethasone (10 nM) in culture medium. Two weeks later, plates were washed, fixed with 4% paraformaldehyde, and stained with 2% alizarin red for visualization of characteristic calcium deposits. Adipogenic differentiation was induced by the addition of dexamethasone (100 nM) and insulin (6 ng/ml) in F12 medium supplemented with 1% fetal calf serum, 1% glutamine, 1% penicillin-streptomycin. Cells were fixed in 4% parafomaldehyde and stained with 0.3% Oil Red solution in 60% isopropanol to assess lipid-laden vacuoles.

Abdi R., Moore R., Sakai S., Donnelly C.B., Mounayar M, & Sackstein R. (2015). HCELL Expression on Murine MSC Licenses Pancreatotropism and Confers Durable Reversal of Autoimmune Diabetes in NOD Mice. Stem cells (Dayton, Ohio), 33(5), 1523-1531.

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Purification and Characterization of T Cell Subsets

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Pooled spleens and lymph nodes were collected from WT or Plac8^-/- mice and processed by gently pressing them through a 70 μm filter. Where noted, CD4 and CD8 T cells were subsequently purified using a mouse T cell negative selection kit (Stemcell Technologies, Vancouver, Canada) according to manufacturer’s instructions. T cells were stained for 15 min at 4°C in PBS + 0.1% BSA (Gemini Bio-Products) using anti-mouse antibodies purchased from eBioscience and Tonbo Biosciences (both San Diego, CA): CD16/CD32 (93), CD4 (IM7), CD8 (53–6.7), CD44 (IM7), CD62L (MGL-14), CD45RB (C363.16A), and CD25 (PC61.5). Propidium iodide (Sigma-Aldrich) was used to exclude dead cells (PI⁺) from the sorted population. For T cell in vitro assays, live cells were designated as CD4⁺ or CD8⁺ T cells before sorting out naïve T cells (CD62L⁺, CD44^lo) or memory T cells (CD44^hi). For the T cell transfer model of colitis, live CD4⁺ T cells were sorted as (CD25^-, CD45RB^hi) effector cells.

Slade C.D., Reagin K.L., Lakshmanan H.G., Klonowski K.D, & Watford W.T. (2020). Placenta-specific 8 limits IFNγ production by CD4 T cells in vitro and promotes establishment of influenza-specific CD8 T cells in vivo. PLoS ONE, 15(7), e0235706.

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TWEAK and TWEAKR Expression in Mouse Aorta

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Protein samples prepared from mouse aortic arch homogenates were electrophoresed and separated on 15% SDS-polyacrylamide gels and transferred to a PVDF membrane (Millipore), which was subsequently blocked with 1% BSA in 0.05% Tween-20 TBS and incubated overnight at 4 °C with the following primary antibodies: anti-TWEAK (1:500, SAB, China) and anti-TWEAKR (1:500, SAB, China). Anti-GAPDH (1:5000, Invitrogen, USA). The membrane was washed and incubated with the appropriate secondary antibodies for 2 h at room temperature; anti-mouse antibodies were from Invitrogen (USA). Protein bands were detected by FluorChem E (Protein Simple, USA).

Zhang K., Kan H., Mao A., Geng L, & Ma X. (2021). Single-cell analysis of salt-induced hypertensive mouse aortae reveals cellular heterogeneity and state changes. Experimental & Molecular Medicine, 53(12), 1866-1876.

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Immunofluorescence Imaging of Transfected Cells

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The acid and alkali solutions were prepared in advance in a 15- mm microscope glass; the NIH3T3 cells or UACC903 cells were placed in 24-well plates. The cells were transfected with 200 ng of each plasmid construct. After transfection for 48 h, the cells were fixed with 4% paraformaldehyde at room temperature for 30 min. Then they were permeabilized for 1 h in PBS/0.2% Triton X-100 (Bio Basic, Amherst, NY, USA), blocked with solution (PBS, 3% bovine serum albumin, 5% goat serum) for 1 h at room temperature and incubated with primary anti-Flag M2 (1:600 dilution) at 4 °C overnight. The cells were washed with PBS/0.2% Triton X-100 for 5 min three times, and incubated for 2 h by avoiding light for preparing fluorescence-labeled secondary anti-mouse antibodies (1:300 dilution; Invitrogen). Then they were incubated with 4,6-diamino-2-phenylindole (DAPI; Invitrogen) for 3 min. Cells were detected by fluorescence images with a laser scanning confocal system installed on a Carl Zeiss microscope (Zeiss, GÖttingen, Germany) with a × 100 oil-immersion objective. Images were analyzed using the Metaphor software package (Zeiss).

Sun J., Hao Z., Luo H., He C., Mei L., Liu Y., Wang X., Niu Z., Chen H., Li J.D, & Feng Y. (2017). Functional analysis of a nonstop mutation in MITF gene identified in a patient with Waardenburg syndrome type 2. Journal of Human Genetics, 62(7), 703-709.

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Antibody-based Investigation of DNA Repair Factors

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Mouse anti-DDX5 (A-5, sc-166167) monoclonal and rabbit anti-RAD51 (H-92, sc-8349) and anti-Ku80 (H-300, sc-9034) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Rabbit anti-53BP1 (NB100-304) was from Novus Biologicals (Littleton, CO). Mouse anti-γH2AX (05-636), anti-p68 (DDX5) (clone204, 05-850) monoclonal antibodies, and rabbit anti-BRCA1 antibody (07-434) were obtained from Millipore (Billerica, MA). Anti-cyclin A (BD611268) monoclonal antibody was from BD Biosciences (San Jose, CA). Anti-BrdU antibody (RPN202) was from Cytiva. V5 mouse mAb antibody was from Life Technologies (#R96025). S9.6 antibody was purified, in house, from the hybridoma (ATCC^® HB-8730). Propidium iodide and the Alexa Fluor-conjugated goat anti-rabbit antibodies and anti-mouse antibodies were from Invitrogen (Carlsbad, CA). Escherichia coli RNase H was purchased from New England Biolabs. Bromo-2′-deoxyuridine (BrdU), 4-hydroxytamoxifen (4-OHT), protein A Sepharose, mouse anti-Flag, and α-tubulin monoclonal antibodies were from Millipore-Sigma (St Louis, MO). For DRIP, Pierce™ Protein A/G UltraLink™ Resin was purchased from Thermo Fisher (#53133). Shield 1 was purchased from Clontech Laboratories (Mountain View, CA). Protease inhibitor cocktail and protein phosphatase inhibitor cocktail were purchased from Roche (Mississauga, ON, Canada).

Yu Z., Mersaoui S.Y., Guitton-Sert L., Coulombe Y., Song J., Masson J.Y, & Richard S. (2020). DDX5 resolves R-loops at DNA double-strand breaks to promote DNA repair and avoid chromosomal deletions. NAR Cancer, 2(3), zcaa028.

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