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Rabbit anti nanog

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Rabbit anti-NANOG is a primary antibody that specifically recognizes the NANOG protein, a transcription factor that plays a crucial role in the maintenance of embryonic stem cell pluripotency and self-renewal. This antibody can be used in various immunoassay techniques to detect and quantify NANOG expression in samples.

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Lab products found in correlation

Rabbit anti oct3 4, by Santa Cruz Biotechnology (6 mentions) Mouse anti tra 1 60, by Merck Group (4 mentions) Matrigel coated 8 well chamber glasses, by Thermo Fisher Scientific (4 mentions) Mouse anti ssea4, by R&D Systems (4 mentions) Hoechst 33342, by Thermo Fisher Scientific (4 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (4 mentions) Fluorescent mouse es ips cell characterization kit, by Merck Group (2 mentions) Axiocam camera, by Zeiss (2 mentions) Hoechst 33258, by Thermo Fisher Scientific (2 mentions) Rabbit anti gfap, by Merck Group (2 mentions) Goat anti dcx, by Santa Cruz Biotechnology (2 mentions) Mouse anti blbp, by Abcam (2 mentions) Rabbit anti arp3, by Santa Cruz Biotechnology (2 mentions) Rabbit anti eif3d, by Fortis Life Sciences (2 mentions) Abc elite peroxidase staining kit, by Thermo Fisher Scientific (2 mentions) Mouse anti tra 1 81, by Merck Group (2 mentions) Mouse anti gapdh, by Merck Group (2 mentions) Mouse anti βiii tubulin tuj1, by Merck Group (2 mentions) Mouse anti nestin, by Merck Group (2 mentions) Mouse anti map2, by Merck Group (2 mentions)

Spelling variants of this product

Nanog (81 citations) Anti-Nanog (37 citations) Rabbit anti-TM (2 citations)

10 protocols using rabbit anti nanog

1

Immunofluorescent Staining of iPSC Markers

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Immunofluorescent staining was used to examine stem cell markers in iPSC colonies [50 (link)]. Briefly, cells were fixed by 4% paraformaldehyde/PBS for 10–15 min and rinsed with PBS, then permeabilized and blocked with 0.1% Triton X-100/PBS containing 3% BSA for 30 min. After incubation with primary antibodies for 1 h at room temperature or overnight at 4C, the samples were washed three times with PBS and then incubated with secondary antibody for 1 h. The following antibodies were used in the immunostaining: rabbit anti-NANOG (1:100 dilution, Santa Cruz) and rabbit anti-OCT4 (1:100 dilution, Millipore). The cell samples were subsequently incubated with Cy3- or Alexa Fluor 488-labeled secondary antibodies for 1 h. After washing three times with PBS, samples were counterstained with Hoechst 33258 (Invitrogen). Alternatively, the pluripotency of stem cells was examined by Fluorescent Mouse ES/iPS Cell Characterization kit (Cat.#SCR077, Millipore, MA) following the protocol provided by the manufacturer. Fluorescence images were acquired with a Zeiss AxioCam Camera.
Du Z., Wen X., Wang Y., Jia L., Zhang S., Liu Y., Zhou L., Li H., Yang W., Wang C., Chen J., Hao Y., Chen H., Li D., Chen N., Celik I., Zhu Y., Yan Z., Fu C., Liu S., Jiao B., Wang Z., Zhang H., Gülsoy G., Luo J., Qin B., Gao S., Kapranov P., Esteban M.A., Zhang S., Li W., Ay F., Chen R., Hoffman A.R., Cui J, & Hu J.F. (2021). Chromatin lncRNA Platr10 controls stem cell pluripotency by coordinating an intrachromosomal regulatory network. Genome Biology, 22, 233.
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2

iPSC Pluripotency Marker Immunostaining

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Immunofluorescent staining was used to examine stem cell markers in iPSC colonies15 (link). Cells were briefly rinsed by freshly prepared PBS, fixed by freshly made 4% paraformaldehyde for 10min at RT, washed three times in PBS for 5 min each, permeabilized with freshly made 0.5% v/v Triton X-100/PBS on ice for 5min, then blocked in 1% w/v BSA for 30min at RT. After incubation with primary antibodies diluted in 1% BSA (2ug/ml 1:500) for 1-3h at RT, the samples were washed three times in PBS for 5 min each, incubated with secondary antibody for 1h at RT. The following antibodies were used in the immunostaining: rabbit anti-NANOG (1:100 dilution, Santa Cruz). The cell samples were subsequently incubated with Cy3 or Alexa Fluor 488 labeled secondary antibodies for 1 hour. After washing three times with PBS, samples were counterstained with Hoechst 33258 (Invitrogen). Alternatively, the pluripotency of stem cells was examined by Fluorescent Mouse ES/iPS Cell Characterization kit (Cat.#SCR077, Millipore, MA) following the protocol provided by the manufacturer. Fluorescence images were acquired with a Zeiss AxioCam Camera.
Wang C., Jia L., Wang Y., Du Z., Zhou L., Wen X., Li H., Zhang S., Chen H., Chen N., Chen J., Zhu Y., Nie Y., Celic I., Gao S., Zhang S., Hoffman A.R., Li W., Hu J.F, & Cui J. (2020). Genome-wide interaction target profiling reveals a novel Peblr20-eRNA activation pathway to control stem cell pluripotency. Theranostics, 10(1), 353-370.
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3

Immunocytochemistry and Western Blot Protocols

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Primary antibodies used for immunocytochemistry or Western blot include mouse anti-GAPDH (Millipore), mouse anti-Nestin (Millipore), mouse anti-Tuj1/βIII-tubulin (Millipore), mouse anti-Tuj1/βIII-tubulin (Santa Cruz Biotenchnology), mouse anti-TRA-1-60 (Millipore), mouse anti-TRA-1-81 (Millipore), goat anti-Lin28a (R&D Systems), rabbit anti-Nanog (Santa Cruz Biotechnology), rabbit anti-Oct3/4 (Santa Cruz Biotechnology), rabbit anti-DCX44 (link), goat anti-DCX (Santa Cruz Biotechnology), goat anti-SOX2 (Santa Cruz Biotechnology), rabbit anti-PAX6 (Covance), mouse anti-MAP2 (Millipore), mouse anti-BLBP (Abcam), rabbit anti-GFAP (Sigma), rabbit anti-OLIG2 (Abcam), mouse anti-β-Catenin (BD), rabbit anti-Arp3 (Santa Cruz Biotechnology), mouse anti-β-actin (Sigma), rabbit anti-EIF3D (Bethyl Labs), rat anti-αN-catenin obtained from the NIH NICHD Developmental Studies Hybridoma Bank (Catalog: NCAT2, deposited by Masatoshi Takeichi and Shinji Hirano). Fluorophore conjugated secondary antibodies were purchased from Invitrogen, and immunohistochemistry was performed with ABC Elite peroxidase staining kit, using ImmpactDAB as a substrate.
Schaffer A.E., Breuss M.W., Caglayan A.O., Al-Sanaa N., Al-Abdulwahed H.Y., Kaymakçalan H., Yılmaz C., Zaki M.S., Rosti R.O., Copeland B., Baek S.T., Musaev D., Scott E.C., Ben-Omran T., Kariminejad A., Kayserili H., Mojahedi F., Kara M., Cai N., Silhavy J.L., Elsharif S., Fenercioglu E., Barshop B.A., Kara B., Wang R., Stanley V., James K.N., Nachnani R., Kalur A., Megahed H., Incecik F., Danda S., Alanay Y., Faqeih E., Melikishvili G., Mansour L., Miller I., Sukhudyan B., Chelly J., Dobyns W.B., Bilguvar K., Jamra R.A., Gunel M, & Gleeson J.G. (2018). Bi-allelic loss of human CTNNA2, encoding αN-catenin, leads to ARP2/3 over-activity and disordered cortical neuronal migration. Nature genetics, 50(8), 1093-1101.
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4

Immunofluorescence Staining of Human iPSCs

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Human iPSC colonies grown in Matrigel-coated 8-well chamber glasses (Thermo Scientific) were fixed using 4% paraformaldehyde (PFA) and permeabilized with 0.5% Triton X-100. After blocking samples with 5% goat serum in PBST (PBS with 0.1% Tween20), cells were stained with mouse anti-SSEA4 (R&D systems), rabbit anti-OCT3/4 (Santa Cruz Biotechnology), rabbit anti-NANOG (Santa Cruz Biotechnology), and mouse anti-SOX2 (R&D systems). Cells were then incubated with Alexa Fluor-conjugated secondary antibodies (Life Technologies) and Hoechst 33342 (Life Technologies) to visualize the specific stains. Image acquisition was performed on an Eclipse 80i fluorescence microscope (Nikon Instruments).
Lee J., Termglinchan V., Diecke S., Itzhaki I., Lam C.K., Garg P., Lau E., Greenhaw M., Seeger T., Wu H., Zhang J.Z., Chen X., Gil I.P., Ameen M., Sallam K., Rhee J.W., Churko J., Chaudhary R., Chour T., Wang P.J., Snyder M.P., Chang H.Y., Karakikes I, & Wu J.C. (2019). Activation of PDGF Pathway Links LMNA Mutation to Dilated Cardiomyopathy. Nature, 572(7769), 335-340.
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5

Immunofluorescence Staining of Cultured Cells

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The cultured cells were fixed in 4% paraformaldehyde for 20 min at room temperature, subsequently washed with PBS and treated with 0.3% Triton X-100(Sigma) in PBS for 15 min. Then the samples were incubated with PBS containing 2% bovine serum albumin (BSA) at 37°C for 10 min. Incubation with the primary antibodies(mouse anti-c-kit, Santa Cruz, 1:200; Rabbit anti-vimentin, Abcam, 1:100; Rabbit anti-CD34, Bioss, 1:300; Rabbit anti-Nanog, Santa Cruz, 1:100; Rabbit anti-sca-1, Millipore,1:200) was performed at 4°C overnight. The samples were subsequently incubated FITC- or Cy3-conjugated secondary antibodies (Beyotime). The nuclei were counterstained with 1μg /ml DAPI (Roche). Negative controls were obtained by following all the same protocol but the primary antibodies. All experiments were performed in triplicate.
Chang Y., Li C., Gan L., Li H, & Guo Z. (2015). Telocytes in the Spleen. PLoS ONE, 10(9), e0138851.
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6

Immunocytochemical Characterization of Pluripotent Stem Cells

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Human ESC and iPSC colonies grown in Matrigel-coated 8-well chamber glasses (Thermo Scientific) were fixed using 4% paraformaldehyde (PFA) and permeabilized with 0.5% Triton X-100. After blocking samples with 5% goat serum in PBST (PBS with 0.1% Tween20), cells were stained with mouse anti-SSEA4 (R&D systems), rabbit anti-OCT3/4 (Santa Cruz Biotechnology), rabbit anti-NANOG (Santa Cruz Biotechnology), and mouse anti-TRA1-60 (EMD Millipore) antibodies. Cells were then incubated with AlexaFluor-conjugated secondary antibodies (Life Technologies) and Hoechst 33342 (Life Technologies) to visualize the specific stains. Image acquisition was performed on an Eclipse 80i fluorescent microscope (Nikon Instruments).
Kodo K., Ong S.G., Jahanbani F., Termglinchan V., Hirono K., InanlooRahatloo K., Ebert A.D., Shukla P., Abilez O.J., Churko J.M., Karakikes I., Jung G., Ichida F., Wu S.M., Snyder M.P., Bernstein D, & Wu J.C. (2016). iPSC-derived cardiomyocytes reveal abnormal TGFβ signaling in left ventricular non-compaction cardiomyopathy. Nature cell biology, 18(10), 1031-1042.
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7

Pluripotency Marker Immunofluorescence in hESCs

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hESCs were fixed with 4% paraformaldehyde for 20 minutes and then permeabilized with 0.1% Triton X-100 in PBS for 5 min. After a treatment with 5% normal goat serum for 30 min, the cells were incubated with primary antibodies against one of the pluripotency markers mouse anti-Oct-4 (Santa Cruz Biotechnology Inc., CA, 1:250) and rabbit anti-Nanog (Santa Cruz Biotechnology Inc, 1:250), for 18 h at 4°C. The cells were then washed 3 times with PBS and incubated with Alexa Fluor 488-conjugated secondary antibodies (Invitrogen, NY, 1:1000) for 1 h. After washing 3 times with PBS, DAPI (Invitrogen) staining was performed for 5 minutes. The samples were immediately imaged using epi-fluorescence microscopes (Nikon TE 2000-U, Nikon, Japan).
Kim C.Y., Hwang I.K., Kang C., Chung E.B., Jung C.R., Oh H., Jeong Y.H., Moon S.H., Kim J.S., Hong K.S., Park J.H, & Chung H.M. (2018). Improved Transfection Efficiency and Metabolic Activity in Human Embryonic Stem Cell Using Non-Enzymatic Method. International Journal of Stem Cells, 11(2), 149-156.
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8

Immunofluorescence Staining of Human iPSCs

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Human iPSC colonies grown in Matrigel-coated 8-well chamber glasses (Thermo Scientific) were fixed using 4% paraformaldehyde (PFA) and permeabilized with 0.5% Triton X-100. After blocking samples with 5% goat serum in PBST (PBS with 0.1% Tween20), cells were stained with mouse anti-SSEA4 (R&D systems), rabbit anti-OCT3/4 (Santa Cruz Biotechnology), rabbit anti-NANOG (Santa Cruz Biotechnology), and mouse anti-SOX2 (R&D systems). Cells were then incubated with Alexa Fluor-conjugated secondary antibodies (Life Technologies) and Hoechst 33342 (Life Technologies) to visualize the specific stains. Image acquisition was performed on an Eclipse 80i fluorescence microscope (Nikon Instruments).
Lee J., Termglinchan V., Diecke S., Itzhaki I., Lam C.K., Garg P., Lau E., Greenhaw M., Seeger T., Wu H., Zhang J.Z., Chen X., Gil I.P., Ameen M., Sallam K., Rhee J.W., Churko J., Chaudhary R., Chour T., Wang P.J., Snyder M.P., Chang H.Y., Karakikes I, & Wu J.C. (2019). Activation of PDGF Pathway Links LMNA Mutation to Dilated Cardiomyopathy. Nature, 572(7769), 335-340.
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9

Immunocytochemistry and Western Blot Protocols

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Primary antibodies used for immunocytochemistry or Western blot include mouse anti-GAPDH (Millipore), mouse anti-Nestin (Millipore), mouse anti-Tuj1/βIII-tubulin (Millipore), mouse anti-Tuj1/βIII-tubulin (Santa Cruz Biotenchnology), mouse anti-TRA-1-60 (Millipore), mouse anti-TRA-1-81 (Millipore), goat anti-Lin28a (R&D Systems), rabbit anti-Nanog (Santa Cruz Biotechnology), rabbit anti-Oct3/4 (Santa Cruz Biotechnology), rabbit anti-DCX44 (link), goat anti-DCX (Santa Cruz Biotechnology), goat anti-SOX2 (Santa Cruz Biotechnology), rabbit anti-PAX6 (Covance), mouse anti-MAP2 (Millipore), mouse anti-BLBP (Abcam), rabbit anti-GFAP (Sigma), rabbit anti-OLIG2 (Abcam), mouse anti-β-Catenin (BD), rabbit anti-Arp3 (Santa Cruz Biotechnology), mouse anti-β-actin (Sigma), rabbit anti-EIF3D (Bethyl Labs), rat anti-αN-catenin obtained from the NIH NICHD Developmental Studies Hybridoma Bank (Catalog: NCAT2, deposited by Masatoshi Takeichi and Shinji Hirano). Fluorophore conjugated secondary antibodies were purchased from Invitrogen, and immunohistochemistry was performed with ABC Elite peroxidase staining kit, using ImmpactDAB as a substrate.
Schaffer A.E., Breuss M.W., Caglayan A.O., Al-Sanaa N., Al-Abdulwahed H.Y., Kaymakçalan H., Yılmaz C., Zaki M.S., Rosti R.O., Copeland B., Baek S.T., Musaev D., Scott E.C., Ben-Omran T., Kariminejad A., Kayserili H., Mojahedi F., Kara M., Cai N., Silhavy J.L., Elsharif S., Fenercioglu E., Barshop B.A., Kara B., Wang R., Stanley V., James K.N., Nachnani R., Kalur A., Megahed H., Incecik F., Danda S., Alanay Y., Faqeih E., Melikishvili G., Mansour L., Miller I., Sukhudyan B., Chelly J., Dobyns W.B., Bilguvar K., Jamra R.A., Gunel M, & Gleeson J.G. (2018). Bi-allelic loss of human CTNNA2, encoding αN-catenin, leads to ARP2/3 over-activity and disordered cortical neuronal migration. Nature genetics, 50(8), 1093-1101.
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10

Immunocytochemical Characterization of Pluripotent Stem Cells

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Human ESC and iPSC colonies grown in Matrigel-coated 8-well chamber glasses (Thermo Scientific) were fixed using 4% paraformaldehyde (PFA) and permeabilized with 0.5% Triton X-100. After blocking samples with 5% goat serum in PBST (PBS with 0.1% Tween20), cells were stained with mouse anti-SSEA4 (R&D systems), rabbit anti-OCT3/4 (Santa Cruz Biotechnology), rabbit anti-NANOG (Santa Cruz Biotechnology), and mouse anti-TRA1-60 (EMD Millipore) antibodies. Cells were then incubated with AlexaFluor-conjugated secondary antibodies (Life Technologies) and Hoechst 33342 (Life Technologies) to visualize the specific stains. Image acquisition was performed on an Eclipse 80i fluorescent microscope (Nikon Instruments).
Kodo K., Ong S.G., Jahanbani F., Termglinchan V., Hirono K., InanlooRahatloo K., Ebert A.D., Shukla P., Abilez O.J., Churko J.M., Karakikes I., Jung G., Ichida F., Wu S.M., Snyder M.P., Bernstein D, & Wu J.C. (2016). iPSC-derived cardiomyocytes reveal abnormal TGFβ signaling in left ventricular non-compaction cardiomyopathy. Nature cell biology, 18(10), 1031-1042.
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