Click it system

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Click-IT™ system is a set of reagents and protocols designed for the detection and analysis of DNA synthesis and cell proliferation. The core function of the Click-IT™ system is to enable the incorporation of modified nucleotides into newly synthesized DNA, which can then be detected and visualized using various detection methods.

Automatically generated - may contain errors

Lab products found in correlation

Canto 2, by BD (1 mentions) Donkey anti rabbit igg cy5, by Jackson ImmunoResearch (1 mentions) Vectashield antifade, by Vector Laboratories (1 mentions) Cy3 donkey anti mouse igg, by Jackson ImmunoResearch (1 mentions) 5 ethynyl 2 deoxyuridine edu, by Thermo Fisher Scientific (1 mentions) Ab6556, by Merck Group (1 mentions) Sab2702214, by Merck Group (1 mentions) Anti laminin, by Merck Group (1 mentions)

Close spelling variants of this product

Click-iT O-GlcNAc Enzymatic Labeling System Click-iT EdU Imaging System Click-IT EdU system Click iT Alexa Fluor Azide system

5 protocols using click it system

Quantifying Cell Proliferation with EdU

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

The nucleoside analog, 5-ethynyl-2-deoxyuridine (EdU), was injected intraperitoneally (25 μg/g body weight) 4 hours before tissue harvesting. Incorporated EdU was detected using the Click-IT™ system (Invitrogen) and analyzed by flow cytometry (BD Canto II).

Chen K.H., Chen C., Lopez T., Radecki K.C., Bustamante K., Lorenson M.Y., Ge X, & Walker A.M. (2018). Use of a novel camelid-inspired human antibody demonstrates the importance of MMP-14 to cancer stem cell function in the metastatic process. Oncotarget, 9(50), 29431-29444.

+ Open protocol

+ Expand

Visualizing DNA Replication and Epigenetic Marks

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

For the visualization of replicating DNA and epigenetic marks of heterochromatic blocks, female Microtus cabrerae cells were transiently transfected with CFP-PCNA and were pulse labeled for 20 min with 10 µM 5′-ethynyl-2′-deoxyuridine (EdU) (Invitrogen, Carlsbad, USA, Cat #: C10337) followed by a 1-h chase prior fixation with 4% paraformaldehyde. EdU was detected with the ClickIT system (Invitrogen) and AlexaFluor 488 followed by an immunostaining with rabbit anti-H3K27me3 (1/200, Upstate, Lake Placid, USA, Cat #: 07-449) and with mouse anti-H3K9me3 (1/100, Active Motif, Carlsbad, USA, Cat #: 39285). The following secondary antibodies were used: donkey anti-mouse IgG-Cy3 (1/200, The Jackson Laboratory, Bar Harbour, USA, Cat #: 715-166-151) and donkey anti-rabbit IgG-Cy5 (1/200, The Jackson Laboratory, Bar Harbor, USA, Cat #: 711-175-152). DNA was counterstained with 1 µg/ml DAPI for 10 min at RT, and cells were mounted afterward in Vectashield antifade (Vector Laboratories) (Fig. 2).

Heinz K.S., Rapp A., Casas-Delucchi C.S., Lehmkuhl A., Romero-Fernández I., Sánchez A., Krämer O.H., Marchal J.A, & Cardoso M.C. (2019). DNA replication dynamics of vole genome and its epigenetic regulation. Epigenetics & Chromatin, 12, 18.

+ Open protocol

+ Expand

Detecting Dynamic Palmitoylation of Cdc42

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

To determine dynamic palmitoylation, click chemistry using the Click-IT system from Invitrogen according to the manufacturer’s instruction was performed. Therefore, N1E-115 cells were transfected with GFP-tagged Cdc42-palm. 12 h after transfection; the cells were either incubated with 50 μM palmitic acid azide or just dimethyl sulfoxide for additional 4 h. After the cells got lysed, 200 μg of total protein were incubated with 40 μM biotin alkylene in the presence of 2 mM CuSO₄. The cell lysates were further processed by methanol–chloroform precipitation. Additionally, cell lysates were immune-precipitated with polyclonal rabbit GFP antibody and protein A-Sepharose. After washing, the protein complex was eluted, purified by trichloroacetic acid-acetone precipitation, and lysates were subjected to SDS-PAGE and Western blotting. Former palmitoylated Cdc42-palm was detected using a HRP-conjugated Biotin antibody (1:500), whereas total Cdc42-palm was detected using a HRP-conjugated GFP antibody (1:1000).

Wirth A., Labus J., Abdel Galil D., Schill Y., Schmidt S., Bunke T., Gorinski N., Yokoi N., Fukata M, & Ponimaskin E. (2022). Palmitoylation of the small GTPase Cdc42 by DHHC5 modulates spine formation and gene transcription. The Journal of Biological Chemistry, 298(6), 102048.

+ Open protocol

+ Expand

Avian Embryo Labeling and Staining

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

Chick and emu embryos were windowed at HH18 and 100μL of 20mM EDU was injected between the embryo and the yolk. Embryos were incubated for 30 minutes following injection and then fixed in 4% formaldehyde and imbedded in 7.5% gelatin and 7.5% sucrose. EDU was visualized using the Click-It® system (ThermoFisher). Anti-laminin (Sigma L9393) was used at 1:50, phalloidin (ThermoFisher A12381) was used at 1:100, anti-Pax3 (R&D Systems AF2457) was used at 1:50), anti-GFP (Abcam Ab6556) and anti-RFP (Sigma SAB2702214) were used at 1:500.

Young J.J., Grayson P., Edwards S.V, & Tabin C.J. (2019). Attenuated Fgf signaling underlies the forelimb heterochrony in the emu Dromaius novaehollandiae. Current biology : CB, 29(21), 3681-3691.e5.

+ Open protocol

+ Expand

In Vivo Cell Proliferation Assay

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

For in vivo proliferation studies, mice were injected intraperitoneally with 1 mg of 5-(ethynyl-2′-deoxyuridine) (EdU) 24 hours prior to sacrifice. In preparation for subsequent flow cytometry analysis, cell populations were immunostained as described above, fixed in 4% paraformaldehyde (PFA) and further treated for EdU staining following the manufacturer’s instructions of the Click-iT System (ThermoScientific). AlexaFluor647-conjugated azide was used to detect incorporated EdU. Stained cells were analyzed by LSRII using the BD Diva software and analyzed by FlowJo. Proliferation was quantified and expressed as percentage of EdU-positive cells.

Christ A., Günther P., Lauterbach M.A., Duewell P., Biswas D., Pelka K., Scholz C.J., Oosting M., Haendler K., Baßler K., Klee K., Schulte-Schrepping J., Ulas T., Moorlag S.J., Kumar V., Park M.H., Joosten L.A., Groh L.A., Riksen N.P., Espevik T., Schlitzer A., Li Y., Fitzgerald M.L., Netea M.G., Schultze J.L, & Latz E. (2018). Western Diet Triggers NLRP3-Dependent Innate Immune Reprogramming. Cell, 172(1-2), 162-175.e14.

+ 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!