Alexa 647

Manufactured by Cell Signaling Technology

Sourced in United States

Alexa Fluor® 647 is a fluorescent dye with maximum excitation and emission wavelengths of 650 nm and 665 nm, respectively. It is commonly used for labeling and detection applications in various life science research techniques.

Automatically generated - may contain errors

Lab products found in correlation

Alexa 488, by Cell Signaling Technology (6 mentions) β actin, by Abcam (2 mentions) Mapk4, by Proteintech (2 mentions) F4 80, by Abcam (2 mentions) Nanozoomer 2.0 rs, by Hamamatsu Photonics (2 mentions) Dm4000 upright fluorescence microscope, by Leica (2 mentions) A7811 clone ea 53, by Merck Group (2 mentions) Cleaved parp, by Merck Group (1 mentions) Rpa32, by Abcam (1 mentions) Dna pkcs, by Abcam (1 mentions) Dna pkcs s2056, by Abcam (1 mentions) Atm s1981, by Abcam (1 mentions) Rpa32 s4 8, by Fortis Life Sciences (1 mentions) Rpa32 s33, by Fortis Life Sciences (1 mentions) Alexa fluor 647, by Cell Signaling Technology (1 mentions) Facsaria, by BD (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Alexa fluor 488, by Merck Group (1 mentions) Anti dsrna j2, by Techcomp Instruments (1 mentions) Prolong gold antifade mountant, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Alexa Fluor® 647 Conjugate (24 citations) Alexa Fluor 647 phalloidin (14 citations) Alexa Fluor 647-conjugated anti-rabbit IgG (6 citations) Anti-rabbit IgG Alexa Fluor 647 (4 citations) Alexa Fluor® 647 conjugated anti-rat IgG (3 citations) Alexa Fluor 647-conjugated anti-mouse IgG (3 citations) Alexa-Fluor-647-conjugated rabbit anti-cleaved caspase-3 (9602S) antibody (2 citations) Alexa Fluor 647 anti-mouse antibodies (2 citations) Alexa Fluor 647-conjugated anti-rabbit IgG (4414) (2 citations) Anti-HA-Tag (Alexa Fluor® 647 Conjugate) (2 citations)

14 protocols using alexa 647

Comprehensive Antibody Panel for DNA Damage Response

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary antibodies used were from Cell Signalling unless otherwise mentioned: β-actin (Abcam #ab6276), cleaved PARP (#5625), CDK1 Y15 (#9111), CHK1 (#2360), CHK1 S345 (#2348), H2AX (#7631), H2AX S139 (Millipore #05-636), H3 (#9715), H3 S10 (#3377), RRM2 (ABNOVA #H00006241-M01), ATR (Santa Cruz #1887), ATR T1989 (Gene Tex #GTX128145), CHK2 (#2662), CHK2 T68 (Abcam #3501), KAP1 (Abcam #10484), KAP1 S824 (Abcam #133440), DNA-PKcs (Abcam #70250), DNA-PKcs S2056 (Abcam #18192), ATM (Abcam #78), ATM S1981 (Abcam #81292), RPA32 (Abcam #2175), RPA32 S4/8 (Bethyl Laboratories #A300-245A), RPA32 S33 (Bethyl Laboratories #A300-246A).
For secondary antibodies, Alexa 488 (#4408, #4412) and Alexa 647 (#4410, #4414) from Cell Signalling were used in immunostaining. IRDye800-conjugated (#925-32210, #926-33210) and IR680-conjugated (#926-68070, #926-68021) antibodies from LICOR were used in immunoblotting.

Wallez Y., Dunlop C.R., Johnson T.I., Koh S.B., Fornari C., Yates J.W., Bernaldo de Quirós Fernández S., Lau A., Richards F.M, & Jodrell D.I. (2018). The ATR inhibitor AZD6738 synergizes with gemcitabine in vitro and in vivo to induce pancreatic ductal adenocarcinoma regression. Molecular cancer therapeutics, 17(8), 1670-1682.

+ Open protocol

+ Expand

Phospho-signaling in Bone Marrow Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Bone marrow cells were starved for 1 hr in IMDM 2% FBS at 37 °C, stained with lineage antibodies, followed by staining with fluorochrome-conjugated antibodies against surface markers, as described above. Post-staining cells were stimulated with 100 ng/ml mSCF (Peprotech #250–03) in 2% PBS-FBS for 5 min at 37 °C. Stained and stimulated cells were fixed and permeabilized with Cytofix/Cytoperm as described above and stained with phospho-S6 (Ser235/236) - Alexa 488 (Cell Signaling Technology, 4803 S) (1:100) and phospho-AKT (Ser473) - Alexa647 (Cell Signaling Technology, 2337 S), phospho-4E-BP1 (Thr37/46) - Alexa Fluor647 -(Cell Signaling Technology, 5123 S) of pERK1(T202/Y204) - Alexa 488 (Cell Signaling 4374) at 1:20 dilutions. Cells were washed with Perm/Wash buffer to remove residual and unbound antibodies, and resuspended in fresh Perm/Wash buffer, followed by flow cytometry analysis on the Cytek Aurora. Analysis of all flow cytometry data was performed using FlowJo software (v9, v10).

Folgado-Marco V., Ames K., Chuen J., Gritsman K, & Baker N.E. (2023). Haploinsufficiency of the essential gene Rps12 causes defects in erythropoiesis and hematopoietic stem cell maintenance. eLife, 12, e69322.

+ Open protocol

+ Expand

Isolation and Characterization of Neuronal and Glial Nuclei

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fresh frozen dlPFC and hippocampus samples were retrieved from -80°C storage and thawed on ice, then disrupted with a handheld homogenizer. Samples were fixed with 1% paraformaldehyde for 10 min at room temperature. Fixation was quenched with glycine for 5 min. Nuclei were isolated by dounce-homogenization followed by filtration through a 70 μM cell strainer (cat no. 21008-952, VWR, Radnor PA). To immunotag cell type specific nuclei, anti-NeuN antibody conjugated to Alexa Fluor 488 (cat no. MAB377X, EMD Millipore, Burlington MA), and anti-PU.1 antibody conjugated to Alexa 647 (cat no. 2240S, Cell Signaling Technology, Danvers MA) were incubated with nuclei at 4°C for 1 h and overnight, respectively. Samples were strained through a 40 μm filter (21008-949, VWR) and stained with the nuclear marker DAPI (D9542, Sigma Aldrich, St. Louis MO) before flow cytometry. First, single nuclei were gated from debris and doublets using DAPI staining. Second, NeuN+ nuclei were gated from NeuN- nuclei. Lastly, NeuN- nuclei were gated as either PU.1+ or PU.1- negative based on average PU.1-647 fluorescence distribution. Fluorescence activated nuclei sorting was performed until 400,000 nuclei were collected for each cell type (NeuN+, Pu.1+, and NeuN-/Pu.1-) using the FACSAria (BD Biosciences, US).

Ramamurthy E., Welch G., Cheng J., Yuan Y., Gunsalus L., Bennett D.A., Tsai L.H, & Pfenning A.R. (2023). Cell type-specific histone acetylation profiling of Alzheimer’s disease subjects and integration with genetics. Frontiers in Molecular Neuroscience, 15, 948456.

+ Open protocol

+ Expand

Immunostaining of dsRNA and dsDNA

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were grown on poly-l-ornithine-pretreated coverslips placed in 12-well plates. Knockout cells were induced by 1 μg ml^–1 doxycycline for 3 d, and compound-treated cells were treated with 1 μM MS023 or DMSO for 5 d. After treatment, cells were washed in PBS and fixed in 4% formaldehyde for 15 min at room temperature. Cells were then washed three times with PBS, permeabilized in 0.5% (vol/vol) Triton X-100 for 10 min and blocked in 5% bovine serum albumin in PBS for 1 h at room temperature. Primary antibody was added (anti-dsRNA J2 (Scicons, 10010200) diluted 1:500 and anti-dsDNA (Abcam, ab27156) diluted 1:1,000) and incubated at 4 °C overnight. Secondary anti-mouse IgG Alexa 647 (Cell Signaling Technology, 4410S) was diluted 1:1,000 and incubated for 1 h in a black container at room temperature. Coverslips were washed with PBS, and DAPI-containing mountant (Invitrogen ProLong Gold Antifade Mountant, P36930) was used. Images were collected on a A1 HD25 Single-Photon confocal microscope and analyzed by NIS-Elements (v5.21.03).

Wu Q., Nie D.Y., Ba-alawi W., Ji Y., Zhang Z., Cruickshank J., Haight J., Ciamponi F.E., Chen J., Duan S., Shen Y., Liu J., Marhon S.A., Mehdipour P., Szewczyk M.M., Dogan-Artun N., Chen W., Zhang L.X., Deblois G., Prinos P., Massirer K.B., Barsyte-Lovejoy D., Jin J., De Carvalho D.D., Haibe-Kains B., Wang X., Cescon D.W., Lupien M, & Arrowsmith C.H. (2022). PRMT inhibition induces a viral mimicry response in triple-negative breast cancer. Nature Chemical Biology, 18(8), 821-830.

+ Open protocol

+ Expand

Molecular Markers for Cell Cycle and Apoptosis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary antibodies used were from Cell Signalling unless otherwise mentioned: β-actin (Abcam #ab6276), BrdU (BD Pharmingen #555627), cleaved caspase 3 (#9664), CDK1 (Abcam #ab18), CDK1 Y15 (#9111), CHK1 (#2360), CHK1 S296 (#2349), CHK1 S345 (#2348), ENT1 (Abcam #ab135756), H2AX (#7631), H2AX S139 (Millipore #05-636), H3 (#9715) and H3 S10 (#3377). For secondary antibodies, Alexa 488 (#4408, #4412) and Alexa 647 (#4410, #4414) from Cell Signalling were used in immunostaining. IRDye800-conjugated (#925-32210, #926-33210) and IR680-conjugated (#926-68070, #926-68021) antibodies from LICOR were used in immunoblotting.

Koh S.B., Wallez Y., Dunlop C.R., de Quirós Fernández S.B., Bapiro T.E., Richards F.M, & Jodrell D.I. (2018). Mechanistic distinctions between CHK1 and WEE1 inhibition guide the scheduling of triple therapy with gemcitabine. Cancer research, 78(11), 3054-3066.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Amyloid-Beta

Check if the same lab product or an alternative is used in the 5 most similar protocols

Frozen hippocampal sections were hydrated, washed with PBS, and repaired with antigen at a high temperature for 15 min. Then the sections were naturally cooled, and endogenous peroxidase blocker was added dropwise. After incubation for 20 min at room temperature, the sections were washed three times with PBS for 5 min each time. Goat serum was added dropwise and blocked at room temperature for 20 min. The serum was removed, the Aβ antibody, 6E10 (1:500, Covance, Princeton, NJ, USA. A diluent was purchased from Beyotime, China, P0103) was added and the sample was incubated overnight in a 4°C refrigerator. The next day, the samples were rewarmed at room temperature and washed with PBS three times, for 5 min each time. The secondary antibody (alexa‐647; 1:100;Cell Signaling Technology; 4414S. A diluent was purchased from Beyotime, China, P0108) was added and incubated at room temperature for 1 hr. PBS was used to rinse the sample three times, for 5 min each time. Finally, an anti‐fluorescence quenching agent was added to seal the slides. Fluorescence microscopy was used to observe and take photographs for statistical analysis.

Xu Y., Hu R., He D., Zhou G., Wu H., Xu C., He B., Wu L., Wang Y., Chang Y., Ma R., Xie M, & Xiao Z. (2020). Bisdemethoxycurcumin inhibits oxidative stress and antagonizes Alzheimer's disease by up‐regulating SIRT1. Brain and Behavior, 10(7), e01655.

+ Open protocol

+ Expand

Yeast-Based Screening of Interferon Variants

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

For both type I and III IFNs, the cytokines were displayed on yeast as previously described (Levin et al., 2012 (link)) but containing a 3C rhinovirus protease tag at the N terminus. Staining and selection was performed via streptavidin-phycoerythrin labeled receptors with separation of the receptor-yeast population by paramagnetic anti-phycoerythrin microbeads (Miltenyi; MACS). Expression on the yeast surface was assayed by staining with the Myc-tag antibody conjugated to Alexa 647 (Cell Signaling). Progression of the enrichment was monitored by the receptor yeast staining by flow cytometry (BD Accuri). A site-directed library was used for engineering type I IFNs. A round of error-prone PCR and DNA shuffling (Brideau-Andersen et al., 2007 (link)) was used for the second-generation of IFN-λ3 variants. Single-point variants of IFN-λ3 were generated by Quick-Change Mutagenesis, transformed, and displayed on yeast.

Mendoza J.L., Schneider W.M., Hoffmann H.H., Vercauteren K., Jude K.M., Xiong A., Moraga I., Horton T.M., Glenn J.S., de Jong Y.P., Rice C.M, & Garcia K.C. (2017). The IFN-λ-IFN-λR1-IL-10Rβ Complex Reveals Structural Features Underlying Type III IFN Functional Plasticity. Immunity, 46(3), 379-392.

+ Open protocol

+ Expand

Cell Doubling and Clonogenic Growth Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

For cell doubling time measurements, cells were plated into six-well dishes in triplicate and counted at three days intervals. For flow cytometry analysis, ethanol-fixed cells were stained with 1 μg/mL Hoechst 33258 (Thermo Fisher) and Phospho Histone H3 (Ser10) antibody conjugated with Alexa647 (Cell Signaling) and analyzed with a SH800 cell sorter (Sony). For confirmation of cell-cycle synchronization, ethanol-fixed cells were stained with 10 μg/mL propidium iodide and 50 μg/mL RNase A and analyzed for DNA content by flow cytometry on a BD LSR II instrument (BD Biosciences). For clonogenic growth assays, 100 cells were plated into six-well dishes in triplicate for 21 days. Methanol-fixed colonies were stained with a 0.5% crystal violet, 25% methanol solution, and quantified using ImageJ software.

Kim D.H., Han J.S., Ly P., Ye Q., McMahon M.A., Myung K., Corbett K.D, & Cleveland D.W. (2018). TRIP13 and APC15 drive mitotic exit by turnover of interphase- and unattached kinetochore-produced MCC. Nature Communications, 9, 4354.

+ Open protocol

+ Expand

Immunofluorescence Analysis of MCPIP-1 and RgpA in Gingival Tissue and Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Slides with gingival tissue or with TIGK cells were blocked with PBS containing 5% FBS, 1% BSA, 0.05% Tween 20, and 2 mM EDTA for 1 h at room temperature (RT). Slides were treated with 0.1% saponin in PBS for 30 min at RT and stained with primary antibodies diluted in buffer (3% BSA and 0.1% saponin in PBS). TIGK cells were stained for 1 h at RT with mouse anti-MCPIP-1 (10 μg/ml; R&D) and rabbit anti-RgpA (10 μg/ml) antibodies; gingival tissues were stained with rabbit anti-Mcpip-1 (13.5 μg/ml; GeneTex). Slides were washed with 0.1% saponin in PBS and stained with secondary antibodies for 45 min at RT. All slides were incubated with goat anti-rabbit antibodies conjugated with Alexa 488 (1:500; Cell Signaling); in addition, TIGK cells were stained with goat anti-mouse antibodies conjugated with Alexa 647 (1:500; Cell Signaling). After several washes with 0.1% saponin in PBS, cell nuclei were stained with Hoechst 33342 (1 μg/ml) for 10 min at RT. Then slides were washed in PBS and mounted in fluorescence medium (Dako). Images were captured with a confocal laser scanning microscope (LSM 880; Zeiss). Quantification of fluorescence signal was with ImageJ software and is displayed as corrected total cell fluorescence (CTCF) or as a percentage of area for fluorescence signal.

Gasiorek A., Dobosz E., Potempa B., Ciaston I., Wilamowski M., Oruba Z., Lamont R.J., Jura J., Potempa J, & Koziel J. (2021). Subversion of Lipopolysaccharide Signaling in Gingival Keratinocytes via MCPIP-1 Degradation as a Novel Pathogenic Strategy of Inflammophilic Pathobionts. mBio, 12(3), e00502-21.

+ Open protocol

+ Expand

Immunofluorescence Analysis of Lung Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lung tissues were fixed in 4% paraformaldehyde, embedded in paraffin, and cut into 4 μm-thick sections. Briefly, the slides were incubated with corresponding primary antibodies (MAPK4: Proteintech, USA, 26102–1-AP; F4/80: Abcam, UK, ab60343) and secondary antibody (Alexa 647: Cell Signaling Technology, USA, 4414S) after deparaffinization and rehydration. Then, the slides were counterstained with DAPI (Beyotime, China, C1002) and observed by Olympus microscope.

Mao L., Zhou Y., Chen L., Hu L., Liu S., Zheng W., Zhao J., Guo M., Chen C., He Z, & Xu L. (2020). Identification of atypical mitogen-activated protein kinase MAPK4 as a novel regulator in acute lung injury. Cell & Bioscience, 10, 121.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!