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Anti rbpj

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Anti-RBPJ is a primary antibody used to detect the RBPJ protein in various experimental techniques, such as Western blotting, immunoprecipitation, and immunohistochemistry. RBPJ is a transcription factor that plays a key role in the Notch signaling pathway, which is involved in cell fate determination and tissue development.

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Lab products found in correlation

Anti notch1, by BioLegend (2 mentions) Anti histone h3 trimethylated on lysine 4 h3k4me3, by Abcam (2 mentions) Anti dimethylated on lysine 27 h3k27me2, by Merck Group (2 mentions) Anti flag, by Merck Group (1 mentions) Anti rat igg hrp, by Jackson ImmunoResearch (1 mentions) Anti vinculin, by Abcam (1 mentions) Anti tbp, by Abcam (1 mentions) Anti vp16, by Santa Cruz Biotechnology (1 mentions) Anti mouse igg hrp, by Cell Signaling Technology (1 mentions) Sheep anti mouse igg hrp, by GE Healthcare (1 mentions) Anti gapdh, by Abcam (1 mentions) Anti rabbit igg hrp, by Cell Signaling Technology (1 mentions) Anti sumo2 3, by Abcam (1 mentions) Anti notch1, by Abcam (1 mentions) Chip it express enzymatic kit, by Active Motif (1 mentions) Anti nucleolin, by Abcam (1 mentions) Anti jag1, by Cell Signaling Technology (1 mentions) Anti phospho yap ser127, by Cell Signaling Technology (1 mentions) Anti actin, by Merck Group (1 mentions) Anti oct4, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

Rabbit anti-Rbpj (2 citations) Anti-Rbpj antibody (2 citations)

5 protocols using anti rbpj

1

ChIP-qPCR Analysis of T-reg Cells

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Chromatin immunoprecipitation on purified Treg cells was performed with Agarose ChIP Kit (Pierce) and anti-RBPJ (Cell signaling), anti-Notch1 (biolegend), anti- Foxp3 (MBL), anti-Histone H3 trimethylated on Lysine 4 (H3K4me3) (abcam), anti-dimethylated on Lysine 27 (H3K27me2) (Millipore) and respective isotype control antibodies. Purified DNA was subjected to real-time PCR with primers flanking RBPJ binding site at Ifng CNS-22 or with primers flanking Foxp3 binding sites on Pde3b and Ikzf2 as previously described 40 37.
Charbonnier L.M., Wang S., Georgiev P., Sefik E, & Chatila T.A. (2015). Control of peripheral tolerance by regulatory T cell-intrinsic Notch signaling. Nature immunology, 16(11), 1162-1173.
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2

Comprehensive Antibody Inventory for Research

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The following antibodies were used in this study: anti-L3MBTL2 (Active Motif, 39,569), anti-E2F6 (Diagenode, C15410314), anti-RBPJ (Cell signaling, 5313), rabbit IgG (Diagenode, C15410206), mouse IgG (Santa Cruz, sc2025), anti-Flag (Merck, F3165, F4042), anti-Flag HRP (Merck, A8592), anti-GFP (Merck/Roche, 11,814,460,001), anti-VP16 (Santa Cruz, sc-7545), anti-GAPDH (Abcam, ab8245), anti-TBP (Abcam, ab818), anti-GST (kind gift from Dr. M.L. Schmitz), anti-SUMO2/3 (Abcam, ab81371), anti-VINCULIN (Abcam, ab130007), anti-mouse IgG-HRP (Cell signaling, 7076 or Amersham NXA931), anti-rabbit IgG HRP (Cell signaling, 7074), anti-rat IgG HRP (Jackson ImmunoResearch, 112-035-072), sheep anti-mouse IgG HRP (GE Healthcare, NA931V), anti-Flag (M2) conjugated agarose beads (Merck, A2220), Hemaglutinin (HA) (Covance, MMS-101P).
Sotomska M., Liefke R., Ferrante F., Schwederski H., Oswald F, & Borggrefe T. (2021). SUMOylated non-canonical polycomb PRC1.6 complex as a prerequisite for recruitment of transcription factor RBPJ. Epigenetics & Chromatin, 14, 38.
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3

ChIP-qPCR Analysis of T-reg Cells

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Chromatin immunoprecipitation on purified Treg cells was performed with Agarose ChIP Kit (Pierce) and anti-RBPJ (Cell signaling), anti-Notch1 (biolegend), anti- Foxp3 (MBL), anti-Histone H3 trimethylated on Lysine 4 (H3K4me3) (abcam), anti-dimethylated on Lysine 27 (H3K27me2) (Millipore) and respective isotype control antibodies. Purified DNA was subjected to real-time PCR with primers flanking RBPJ binding site at Ifng CNS-22 or with primers flanking Foxp3 binding sites on Pde3b and Ikzf2 as previously described 40 37.
Charbonnier L.M., Wang S., Georgiev P., Sefik E, & Chatila T.A. (2015). Control of peripheral tolerance by regulatory T cell-intrinsic Notch signaling. Nature immunology, 16(11), 1162-1173.
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4

ChIP-seq of NOTCH1 and RBPJ

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ChIP assay was performed using ChIP-IT Express Enzymatic Kit (Active Motif). Briefly, cells were cross-linked with 1% formaldehyde and then lysed. After enzymatic digestion, samples were immunoprecipitated with anti-NOTCH1 (Abcam 27526; 1:200 dilution) or anti-RBPJ (Cell Signaling 5313, s; 1:200 dilution), or control normal anti-Rabbit IgG (Cell Signaling Technology). DNA was eluted and quantitative PCR was carried out with specific primers to amplify the RBPJ-binding site of the ERBB3 promoter (forward: 5′-GACGGTGCGGCCAGACTCCA-3′ and reverse: 5′-TCTCCCGGGGATTTGGAT-3′).
Zhang T., Guo L., Creighton C.J., Lu Q., Gibbons D.L., Yi E.S., Deng B., Molina J.R., Sun Z., Yang P, & Yang Y. (2016). A genetic cell context-dependent role for ZEB1 in lung cancer. Nature Communications, 7, 12231.
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5

Immunoblotting analysis of stem cell markers

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Immunoblotting was performed as described, using 30–100µg of lysate per sample (7 (link)), and the following antibodies: anti-YAP (Cell Signaling #4912, 1:1000), anti-phospho-YAP Ser127 (Cell Signaling #4911, 1:1000), anti-CD133 (DSHB #HB7, 1:50), anti-OCT4 (Cell Signaling #2750, 1:250–1:500), anti-RBPJ (Cell Signaling #5313, 1:1000), anti-JAG1 (Cell Signaling #2620, 1:1000), anti-Nucleolin (Abcam #ab22758, 1:1000) and anti-Actin (Sigma #A2066, 1:1000 or Sigma #A5441, 1:5000). Densitometry was performed using ImageJ (NIH), and all values were normalized to the loading control.
Slemmons K.K., Crose L.E., Riedel S., Sushnitha M., Belyea B, & Linardic C.M. (2017). A Novel Notch-YAP Circuit Drives Stemness and Tumorigenesis in Embryonal Rhabdomyosarcoma. Molecular cancer research : MCR, 15(12), 1777-1791.
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