Cy2 and cy5 conjugated secondary antibodies

Manufactured by Jackson ImmunoResearch

Cy2- and Cy5-conjugated secondary antibodies are fluorescent-labeled antibodies designed for use in immunodetection techniques. Cy2 is a green-fluorescent dye, while Cy5 is a far-red fluorescent dye. These conjugated secondary antibodies can be used to detect and visualize target proteins in a variety of applications, such as Western blotting, immunohistochemistry, and flow cytometry.

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Lab products found in correlation

Mouse anti th antibody, by Immunostar (3 mentions) Maraviroc, by Selleck Chemicals (2 mentions) Anti cd8 antibody, by Thermo Fisher Scientific (2 mentions) Anti gfap antibody, by Agilent Technologies (2 mentions) Mouse anti inos antibody, by BD (2 mentions) Anti iba1 antibody, by Abcam (2 mentions) Rabbit anti α syn antibody clone mjfr1, by Abcam (2 mentions) Anti cd4 antibody, by Thermo Fisher Scientific (2 mentions) Ix81 fluorescent microscope, by Olympus (2 mentions) Mouse anti map2, by Merck Group (1 mentions) Formaldehyde, by Polysciences (1 mentions) Goat anti iba1 antibody, by Abcam (1 mentions) Anti phospho p65, by Abcam (1 mentions) Rabbit anti gfap antibody, by Agilent Technologies (1 mentions)

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Cy2- and Cy5-conjugated antibodies (8 citations) Cy2 and Cy5 (4 citations)

7 protocols using cy2 and cy5 conjugated secondary antibodies

Immunostaining Protocol for Neurodegeneration

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Example 1

Maraviroc was purchased from Selleck Chemicals (Houston, Tex.). Mouse anti-TH antibody (Immunostar), rabbit anti-α-syn antibody (clone: MJFR1) (Abcam), anti-CD4 antibody (Thermofisher), anti-CD8 antibody (Thermofisher), anti-Iba1 antibody (Abcam), anti-GFAP antibody (Agilent), and mouse anti-iNOS antibody (BD Bioscience) were purchased from different vendors. Cy2- and Cy5-conjugated secondary antibodies were obtained from Jackson Immuno Research Laboratories (West Grove, Pa.).

US11504360B2. Compositions and methods for treating neurological disorders (2022-11-22). Rush University Medical Center [US]. Inventors: Kalipada Pahan [US].

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TRPV4 and Neuronal Cytoskeleton Characterization

Cited 3 times

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Antibodies used in this study include rabbit polyclonal anti-TRPV4 (Lifespan Biosciences, Inc.), mouse anti-TRPV1 antibody (clone N221/17; NeuroMAB), mouse anti-STOP antibody (Cell Signaling Technology), mouse anti-β-tubulin (EMD Millipore), mouse anti-MAP2, rabbit anti-Tau1, rat anti-MBP antibodies (EMD Millipore), and Cy2- and Cy5-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, Inc.). In particular, the anti-TRPV4 antibody (LS-C94498; Lifespan Biosciences) was extensively validated by using Western blots and immunostaining of wild-type and TRPV4-null mice (as well as TRPV4 cDNA; Ryskamp et al., 2011 (link)). The procedure of immunocytochemistry was previously described (Jukkola et al., 2012 (link), 2013 (link)). In brief, neurons were fixed with 4% formaldehyde (10% ultrapure EM grade and methanol free; Polysciences, Inc.) and 4% sucroseq in PBS for 20 min and then stained with specified antibodies under permeabilized conditions in the presence of 0.2% Triton X-100 to label total proteins. To distinguish axons and dendrites of neurons, the anti-MAP2 (a dendritic marker) antibody was used in costaining.

Gu Y., Jukkola P., Wang Q., Esparza T., Zhao Y., Brody D, & Gu C. (2017). Polarity of varicosity initiation in central neuron mechanosensation. The Journal of Cell Biology, 216(7), 2179-2199.

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Immunofluorescence Analysis of Neuronal Proteins

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Immunofluorescence analysis was performed as described earlier (Mondal et al.; Roy et al., 2008 (link)). Briefly, cells cultured in 8-well chamber slides (Lab-Tek II) were fixed with 4% paraformaldehyde for 20 min followed by treatment with cold ethanol (−20 °C) for 5 min and 2 rinses in PBS. Samples were blocked with 3% BSA in PBST for 30 min and incubated in PBST containing 1% BSA and rabbit anti-NR2A (1:100), anti-GluR1 (1:100), anti-PSD95 (1:100) and anti-CREB (1:100). After three washes in PBST (15 min each), slides were further incubated with cy2- and cy5-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, Inc.). For negative controls, a set of culture slides was incubated under similar conditions without the primary antibodies. The samples were mounted and observed under an Olympus IX81 fluorescent microscope. For tissue staining, brains kept in 4% paraformaldehyde were sectioned in cryostat machine with 30 μm thickness followed by the immunostaining as described before (Ghosh et al., 2007 (link)).

Modi K.K., Rangasamy S.B., Dasarathi S., Roy A, & Pahan K. (2016). Cinnamon converts poor learning mice to good learners: Implications for memory improvement. Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology, 11(4), 693-707.

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Immunohistochemistry in MPTP Mouse Model

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GTB was purchased from Spectrum (New Brunswick, NJ). MPTP was procured from Sigma-Aldrich (St. Louis, MO). Mouse anti–TH antibody (ImmunoStar, Hudson, WI), goat anti-Iba1 antibody (Abcam, Cambridge, MA), rabbit anti-GFAP antibody (Agilent Technologies, Santa Clara, CA), mouse anti-iNOS Ab (BD Biosciences, San Jose, CA), anti GTP-p21^Ras (New East Biosciences) and anti-phospho-p65 (Abcam) antibodies were purchased from different vendors. Cy2-and Cy5-conjugated secondary antibodies were obtained from Jackson Immuno-Research Laboratories (West Grove, PA).

Rangasamy S.B., Dutta D., Mondal S., Majumder M., Dasarathy S., Chandra G, & Pahan K. (2022). Protection of dopaminergic neurons in hemiparkinsonian monkeys by flavouring ingredient glyceryl tribenzoate. Neuroimmune Pharmacology and Therapeutics, 1(1), 7-22.

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Immunofluorescence Labeling Protocol

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Cy2- and Cy5-conjugated secondary antibodies were obtained from Jackson Immuno-Research Laboratories (West Grove, PA). Details about primary antibodies are given in Table S1.

Shelver D., Kerby R.L., He Y, & Roberts G.P. (1997). CooA, a CO-sensing transcription factor from Rhodospirillum rubrum, is a CO-binding heme protein. Proceedings of the National Academy of Sciences of the United States of America, 94(21), 11216-11220.

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Immunohistochemistry of Synucleinopathy

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Maraviroc was purchased from Selleck Chemicals (Houston, TX). Mouse anti-TH antibody (Immunostar), mouse anti-human laminin α5 antibody clone # 4B12 (Millipore), rabbit anti-α-syn antibody (clone: MJFR1) (Abcam), anti-CD4 antibody (Thermofisher), anti-CD8 antibody (Thermofisher), anti-Iba1 antibody (Abcam), anti-GFAP antibody (Agilent), and mouse anti-iNOS antibody (BD Bioscience) were purchased from different vendors. Cy2- and Cy5-conjugated secondary antibodies were obtained from Jackson Immuno Research Laboratories (West Grove, PA).

Mondal S., Rangasamy S.B., Roy A., Dasarathi S., Kordower J.H, & Pahan K. (2019). Low-dose maraviroc, an antiretroviral drug, attenuates the infiltration of T cells into the CNS and protects the nigrostriatum in hemiparkinsonian monkeys. Journal of immunology (Baltimore, Md. : 1950).

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Immunofluorescence Analysis of Synaptic Proteins

Cited 1 time

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Immunofluorescence analysis was performed as described earlier [25] (link), [26] (link), [27] (link). Briefly, cells cultured in 8-well chamber slides (Lab-Tek II) were fixed with 4% paraformaldehyde for 20 min followed by treatment with cold ethanol (−20°C) for 5 min and 2 rinses in PBS. Samples were blocked with 3% BSA in PBST for 30 min and incubated in PBST containing 1% BSA and rabbit anti-NR2A (1∶100), anti-GluR1 (1∶100), anti-PSD95 (1∶100) and anti-CREB (1∶100). After three washes in PBST (15 min each), slides were further incubated with cy2- and cy5-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, Inc.). For negative controls, a set of culture slides was incubated under similar conditions without the primary antibodies. The samples were mounted and observed under an Olympus IX81 fluorescent microscope. For tissue staining, brains were kept in 4% paraformaldehyde and 30-µm slices were sectioned in a cryostat followed by immunostaining as described before [28] (link).

Roy A., Modi K.K., Khasnavis S., Ghosh S., Watson R, & Pahan K. (2014). Enhancement of Morphological Plasticity in Hippocampal Neurons by a Physically Modified Saline via Phosphatidylinositol-3 Kinase. PLoS ONE, 9(7), e101883.

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