Alexa fluor 700 anti mouse cd45

Manufactured by BioLegend

Sourced in United States

Alexa Fluor 700 anti-mouse CD45 is a fluorescently labeled antibody that specifically binds to the CD45 protein expressed on the surface of mouse cells. It is designed for use in flow cytometry applications.

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Lab products found in correlation

Apc or fitc anti mouse cd45, by Cytek Biosciences (3 mentions) Cytoflex lx, by Beckman Coulter (3 mentions) Pe anti mouse cd3, by BioLegend (3 mentions) Lsrfortessa, by BD (3 mentions) Pe anti myc, by Cell Signaling Technology (2 mentions) Cytofix cytoperm kit, by BD (2 mentions) Fitc anti mouse cd3, by BioLegend (2 mentions) Cd71 pe, by Thermo Fisher Scientific (1 mentions) Lsrfortessa x 20 cytometer, by BD (1 mentions) Pe anti mouse il 17a, by BioLegend (1 mentions) Live dead fixable violet dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Cell stimulation cocktail, by Thermo Fisher Scientific (1 mentions) Fortessa, by BD (1 mentions) Pe cf594 anti mouse cd3e, by BD (1 mentions) Zombie nir fixable viability kit, by BioLegend (1 mentions) Pacific blue anti mouse cd4, by BioLegend (1 mentions) Pe anti mouse il 4, by BioLegend (1 mentions) Transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions) Alexa fluor 647 anti mouse foxp3, by BioLegend (1 mentions) Fitc anti mouse cd3ε, by BioLegend (1 mentions)

Spelling variants of this product

Anti-CD45 (228 citations) Anti-mouse CD45 (39 citations) Alexa Fluor 700 (29 citations) CD45-Alexa Fluor 700 (23 citations) Anti-CD45 Alexa Fluor 700 (7 citations) Alexa Fluor 700-conjugated anti-mouse CD45 (2 citations) Alexa Fluor 700 anti-mouse CD45 antibody (2 citations)

12 protocols using alexa fluor 700 anti mouse cd45

Flow Cytometry of Mouse Immune Cells

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Cell samples were washed and stained in flow buffer (PBS, 1% FBS, 0.1% sodium azide) for 20–30 min. Antibodies used in the study are as follows: APC or FITC anti-mouse CD45.1 (clone: A20, Tonbo Biosciences), PE anti-myc (Clone: 71D10, Cell Signaling Technology), anti-Muc16t was generated and conjugated in house (9 (link),25 (link)), Alexa Fluor 700 anti-mouse CD45 (Clone: 30-F11, BioLegend), and PE anti-CD3 mouse (clone: UCHT1, BioLegend). Data was collected using a Guava easyCyte HT Flow Cytometer (Luminex, Austin, Texas, USA), a LSRFortessa (BD, Franklin Lakes, NJ, USA), or a Cytoflex LX (Beckman Coulter, Pasadena, CA). Data were analyzed using Flowjo v10.4 software (Flowjo, Ashland, OR, USA).

Bourne C.M., Wallisch P., Dacek M.M., Gardner T.J., Pierre S., Vogt K., Corless B.C., Bah M.A., Romero-Pichardo J.E., Charles A., Kurtz K.G., Tan D.S, & Scheinberg D.A. (2023). Host Interactions with Engineered T-cell Micropharmacies. Cancer Immunology Research, 11(9), 1253-1265.

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Splenic and Bone Marrow Cell Phenotyping

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Splenic and bone marrow cells were diluted in cold FACS buffer (

PBS + 1 % FBS + 0.02 % sodium azide

), divided into

4 \times 10^{4}

cell aliquots, stained with an antibody cocktail of 1:200 APC anti-mouse TER119 (TONBO biosciences, catalog no. 20-5921-U025); 1:1,000 CD71, PE (eBioscience; catalog no. 12-0711-81); and 1:400 Alexa Fluor® 700 anti-mouse CD45 (BioLegend; catalog no. 103128) in the dark for 30 min at 4°C. Cells were analyzed on a BD LSRFortessa™ X-20 cytometer (BD Biosciences), and data were processed using FlowJo software (v10.5.3; Tree Star).

Egusquiza R.J., Ambrosio M.E., Wang S.G., Kay K.M., Zhang C., Lehmler H.J, & Blumberg B. (2020). Evaluating the Role of the Steroid and Xenobiotic Receptor (SXR/PXR) in PCB-153 Metabolism and Protection against Associated Adverse Effects during Perinatal and Chronic Exposure in Mice. Environmental Health Perspectives, 128(4), 047011.

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Flow cytometry protocol for mouse T cell analysis

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Single-cell suspensions were pelleted and resuspended in PBS with 2% FBS containing fluorophore-conjugated antibodies. Cells were initially stained with antibodies targeting cell surface proteins and with Live/dead Fixable Violet Dead Cell Stain Kit (Thermo Scientific, cat: L34964) for 30 min on ice and washed with PBS containing 2% FBS. For intracellular target staining, cells were then fixed and permeabilized using a Cytofix/Cytoperm kit (BD Biosciences, cat: 554714). For intracellular cytokine staining, single-cell suspensions were plated at 4 × 10⁶ cells per well in a 96-well round bottom plate, resuspended in 1× cell stimulation cocktail (plus protein transport inhibitors, eBioscience, cat: 00-4975-03) and incubated at 37 °C for 4 h before antibody staining. Samples were run on a BD Fortessa and analyzed using FlowJo software (Treestar, version 10.6.2).
The following antibodies were used for mouse flow cytometry: Alexa Fluor® 700 anti-mouse CD45 [Clone: 30-F11] (BioLegend, cat: 103128), PE CF594 anti-mouse Cd3e [Clone: 145 2C11] (BD, cat: 562286), PE-Cyanine5 anti-mouse TCR gamma/delta [Clone: GL-3] (Invitrogen, cat: 15-5711-81), PE anti-mouse IL-17A [Clone: TC11-18H10.1] (BioLegend, cat: 506904).

Cai X., Han M., Lou F., Sun Y., Yin Q., Sun L., Wang Z., Li X., Zhou H., Xu Z., Wang H., Deng S., Zheng X., Zhang T., Li Q., Zhou B, & Wang H. (2023). Tenascin C+ papillary fibroblasts facilitate neuro-immune interaction in a mouse model of psoriasis. Nature Communications, 14, 2004.

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Multiparametric Flow Cytometry Analysis

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Cell samples were washed and stained in flow buffer (PBS, 1% FBS, 0.1% sodium azide) for 20 to 30 minutes. Antibodies used in the study are as follows: APC or FITC anti-mouse CD45.1 (clone: A20, Tonbo Biosciences), PE anti-myc (Clone: 71D10, Cell Signaling Technology), anti-Muc16t was generated and conjugated in house (9, 25 (link)), Alexa Fluor 700 anti-mouse CD45 (Clone: 30-F11, BioLegend), and PE anti-CD3 mouse (clone: UCHT1, BioLegend). Data were collected using a Guava easyCyte HT Flow Cytometer (Luminex), a LSRFortessa (BD Biosciences), or a Cytoflex LX (Beckman Coulter). Data were analyzed using Flowjo v10.4 software (Flowjo).

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Multi-parameter Flow Cytometry Analysis of Immune Cells

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The spleen was dissociated into single cells by mashing the tissue through a
70-µm cell strainer. Red blood cells from spleens were lysed using 0.83% (w/v)
of ammonium chloride. For the detection of immune cell subtypes, the following
antibodies were used: Alexa Fluor 647 anti-mouse FOXP3 (Biolegend, 126407),
Alexa Fluor 700 anti-mouse CD45 (Biolegend, 103127), Brilliant Violet 510
anti-mouse CD8a (Biolegend, 100751), Brilliant Violet 650 anti-mouse CD19
(Biolegend, 115541), Brilliant Violet 785 anti-mouse Ly-6C (Biolegend, 128041),
FITC anti-mouse CD3 (Biolegend, 100203), Pacific Blue anti-mouse CD4 (Biolegend,
100427), PE anti-mouse IL-4 (Biolegend, 504103), PC/Cy7 anti-mouse IFNgamma
(Biolegend, 505825), PE/Dazzle 594 anti-mouse IL-17A (Biolegend, 506937), and
PerCP/Cy5.5 anti-mouse CD11b (Biolegend, 101227). Viable cells were gated using
the Zombie NIR Fixable Viability Kit (Biolegend, 423105). For cell surface
staining, cells were incubated with 10% rat serum for 15 min prior to staining
with fluorescently labeled antibodies for 15 min on ice. For intracellular
staining, cells were fixed, permeabilized, and stained using the Transcription
Factor Staining Buffer Set (Thermo Fisher Scientific) according to the
manufacturer’s protocol. Flow cytometry was carried out and analyzed on a BD
LSRFortessa (BD Biosciences).

Bogie J.F., Grajchen E., Wouters E., Broux B., Stinissen P., Van Wijmeersch B, & Hendriks J.J. (2020). CNS delivery of anti-CD52 antibodies modestly reduces disease severity in an animal model for multiple sclerosis. Therapeutic Advances in Chronic Disease, 11, 2040622320947378.

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Flow Cytometry Analysis of Immune Cells

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Cell samples were washed and stained in flow buffer (PBS, 1% FBS, 0.1% sodium azide) for 20–30 min. Antibodies used in the study are as follows: APC or FITC anti-mouse CD45.1 (clone: A20, Tonbo Biosciences), Alexa Fluor 700 anti-mouse CD45 (Clone: 30-F11, BioLegend), PE anti-CD3 mouse monoclonal antibody (clone: UCHT1, BioLegend). Data was collected using a Guava easyCyte HT Flow Cytometer (Luminex, Austin, Texas, USA), a LSRFortessa (BD, Franklin Lakes, NJ, USA), or Cytoflex LX (Beckman Coulter, Pasadena, CA). Data were analyzed using Flowjo v10.4 software (Flowjo, Ashland, OR, USA).

Bourne C.M., Wallisch P., Dacek M., Gardner T., Pierre S., Vogt K., Corless B.C., Bah M.A., Romero Pichardo J., Charles A., Kurtz K.G., Tan D.S, & Scheinberg D.A. (2023). Host-cell Interactions of Engineered T cell Micropharmacies. bioRxiv.

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Quantifying Eosinophils and T Cells in BALF and Lung

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To quantify the counts of eosinophils, cells from BALF or lung tissues were resuspended in 100 µL RPMI 1640 medium containing BV510 Live/Dead (catalog no. 423101, Biolegend), Alexa Fluor®-700 anti-mouse CD45 (catalog no. 103127, clone 30-F11, Biolegend), KIRAVIA Blue 520^TM anti-mouse F4/80 (catalog no. 123161, clone BM8, Biolegend), Brilliant Violet 421^TM anti-mouse CD11c (catalog no. 117329, clone N418, Biolegend), APC/Cyanine7-anti-mouse CD11b (catalog no. 101226, clone M1/70, Biolegend), and PE anti-mouse CD170 (Siglec-F) (catalog no. 155505, clone S17007L, Biolegend) on ice for 30 minutes. To quantify the counts of T lymphocytes, cells from BALF or lung tissues were resuspended in 100 µL RPMI 1640 medium containing BV510 Live/Dead (catalog no. 423101, Biolegend), FITC anti-mouse CD3ε (catalog no.100306, clone 145-2C11, Biolegend) on ice for 30 min. All cells were washed with PBS and were measured on a BD LSRFortessa^TM Flow Cytometer (BD Biosciences). All antibodies were purchased from Biolegend and were diluted at 1:200. A minimum of 5000 events per plot were collected and analyzed using FlowJo V10 software. The numbers presented in the flow cytometry analysis images are percentage-based. All antibodies were validated for the specified application by respective manufacturer.

Wu X., Yu Y., Wang M., Dai D., Yin J., Liu W., Kong D., Tang S., Meng M., Gao T., Zhang Y., Zhou Y., Guan N., Zhao S, & Ye H. (2024). AAV-delivered muscone-induced transgene system for treating chronic diseases in mice via inhalation. Nature Communications, 15, 1122.

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Multiparametric Analysis of Cell Populations

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Rabbit anti-ABCB1 (1:100, 22336-1-AP, Proteintech) and rabbit anti-CX43 (1:100, 26980-1-AP, Proteintech) were used as primary antibodies to stain the cell suspensions, and Alexa Fluor 647 donkey anti-rabbit IgG (H + L) (1:200, A-31573, Thermo Fisher Scientific) was used as the secondary antibody. PE/Cy7-anti-mouse CD31 (1:100, 102418, BioLegend) and Alexa Fluor 700 anti-mouse CD45 antibodies (1:100, 147716, BioLegend) were used to stain the endothelial and immune cells. Dead cells in samples were excluded by propidium iodide (PI) staining in indicated experiments. Samples were analysed using Canto II with BD FACSDiva software v8.0.1, and the results were analysed using FlowJo v10 (BD Bioscience, USA). Gating strategies have been provided in the figures as well as in Supplementary Fig. 10.

Ni C., Lou X., Yao X., Wang L., Wan J., Duan X., Liang J., Zhang K., Yang Y., Zhang L., Sun C., Li Z., Wang M., Zhu L., Lv D, & Qin Z. (2022). ZIP1+ fibroblasts protect lung cancer against chemotherapy via connexin-43 mediated intercellular Zn2+ transfer. Nature Communications, 13, 5919.

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Flow Cytometry Immune Cell Analysis

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Cell sorting was performed using a FACSAria^™ Cell Sorter (BD Biosciences) at CSHL Flow Cytometry Facility. FlowJo X (Tree Star) software was used for experimental analysis.
The following antibodies were used: Alexa Fluor 700 anti-mouse CD45 (#103127; BioLegend), FITC anti-mouse CD45 (#11-0451; Thermo Fisher), APC/Cy7 anti-mouse CD3ε (#100329; BioLegend), PerCP/Cyanine 5.5 anti-mouse CD4 (#100433; BioLegend), Brilliant Violet 510TM anti-mouse CD8a (#100751; BioLegend), Brilliant Violet 605TM anti-mouse/human CD11b (#101257; BioLegend), Alexa Fluor 700 anti-mouse Ly-6G/Ly-6C (Gr-1) (#108421; BioLegend), FITC anti-mouse CD69 (#104505; BioLegend), PE/Cy7 anti-mouse CD152 (#106313; BioLegend); Brilliant Violet 421TM anti-mouse CD274 (#124315; BioLegend), PE/Dazzle 594 anti-mouse CD279 (#109115; BioLegnd), and FITC anti-mouse F4/80 (#123107; BioLegend).

Ferrer M., Mourikis N., Davidson E.E., Kleeman S.O., Zaccaria M., Habel J., Rubino R., Flint T.R., Connell C.M., Lukey M.J., White E.P., Coll A.P., Venkitaraman A.R, & Janowitz T. (2023). Ketogenic diet promotes tumor ferroptosis but induces relative corticosterone deficiency that accelerates cachexia. bioRxiv.

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Preparation and Characterization of Immune Modulators

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Compounds IN1 (CG14600), IN3 (CG22600), and IN4 (CY16600) were prepared as previously reported.²¹, ²², ²³, ²⁴ Stock solutions for these compounds were made using sterile saline (0.90% NaCl). An LPS isolated from E. coli O55:B5 that does not contain sialic acid was purchased from Sigma Aldrich (cat# L2880).²⁵ Anti‐mouse/human CD11b FITC (Clone M1/70), anti‐mouse CD45 Alexa Fluor 700 (Clone 30‐F11), anti‐mouse/human CD45R/B220 PerCP (Clone RA3‐6B2), anti‐mouse 49b PE (Clone DX5), anti‐mouse F4/80 APC (Clone BM8), anti‐mouse Ly‐6G PE/Cyanine 7 (Clone 1A8), anti‐mouse Ly‐6C Brilliant violet 421 (Clone HK1.4), and anti‐mouse CD115 APC/Cyanine 7 (Clone AFS98) were purchased from Biolegends, USA.

Howlader M.A., Demina E.P., Samarani S., Guo T., Caillon A., Ahmad A., Pshezhetsky A.V, & Cairo C.W. (2022). The Janus‐like role of neuraminidase isoenzymes in inflammation. The FASEB Journal, 36(5), e22285.

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