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Cdp star reagent

Manufactured by Roche
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CDP-Star reagent is a chemiluminescent substrate used for the detection of alkaline phosphatase-conjugated molecules in various immunoassay and nucleic acid detection techniques. It generates a stable luminescent signal that can be measured using a luminometer or imaging system.

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Lab products found in correlation

Hybond, by Cytiva (7 mentions) Pvdf membrane, by Cytiva (7 mentions) Pcr dig probe synthesis kit, by Roche (4 mentions) Hybond n membrane, by GE Healthcare (4 mentions) Las 4000, by Cytiva (4 mentions) Positively charged nylon membrane, by Roche (3 mentions) Las 4000 detection system, by GE Healthcare (3 mentions) Northernmax gly sample loading dye, by Thermo Fisher Scientific (3 mentions) Dig northern blot starter kit, by Roche (3 mentions) Semi dry blotting system, by Bio-Rad (3 mentions) Dig easy hyb, by Roche (3 mentions) Dig northern starter kit, by Roche (2 mentions) Anti β actin, by Merck Group (2 mentions) β actin, by Merck Group (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Positively charged nylon membrane, by Cytiva (2 mentions) Digoxigenin labeled lna probes, by Qiagen (2 mentions) Gapdh, by Abmart (2 mentions) Ultrahyb oligo buffer, by Thermo Fisher Scientific (1 mentions) Hybond n nylon membrane, by Cytiva (1 mentions)

Spelling variants of this product

CDP-Star (201 citations) CDP-Star Detection Reagent (3 citations)

29 protocols using cdp star reagent

1

Northern Blot Analysis of RNA

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Total RNA was separated on a 1.2 % (w/v) formaldehyde agarose gel and then blotted onto Hybond N+ nylon membranes (Amersham Biosciences) by semi-dry transfer (Bio-Rad,
Trans-Blot SD Semi-Dry Transfer Cell). DNA probes were ordered from Integrated DNA Technologies (IDT, San Diego, CA) with digoxigenin incorporated at 3′end. For ACTIN we used the RNA probe provided in the DIG Northern Starter Kit (Roche). Membranes were hybridized overnight using ULTRAhyb-Oligo Buffer (Ambion) at 37 or 42 °C with probes. Visualization was done by X-Ray film using CDP-Star reagents (Roche). X-Ray film was scanned and saved as jpeg files. Brightness and contrast was increased by 20 % for ease of visualization.
Rodríguez-Malavé N.I., Fernando T.R., Patel P.C., Contreras J.R., Palanichamy J.K., Tran T.M., Anguiano J., Davoren M.J., Alberti M.O., Pioli K.T., Sandoval S., Crooks G.M, & Rao D.S. (2015). BALR-6 regulates cell growth and cell survival in B-lymphoblastic leukemia. Molecular Cancer, 14, 214.
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2

NF-κB Expression Analysis in NOZ Cells

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Nucleoprotein was extracted from NOZ cells for measurement of the expression of NF-κB, separated with 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and electrophoretically transferred to polyvinylidene fluoride (PVDF) membranes. The membranes were incubated with monoclonal anti- NF-κB p65 (1:1000; Abcam, Cambridge, UK), with Histone H3 as the internal protein control. Proteins were detected by addition of alkaline phosphatase (AP)-conjugated secondary antibody. Immunoreactive proteins were visualized by addition of CDP STAR reagents (Roche Diagnostics, Germany). Fragment analysis was conducted using Phoretix 1D software.
Du Q., Jiang L., Wang X., Wang M., She F, & Chen Y. (2014). Tumor necrosis factor-α promotes the lymphangiogenesis of gallbladder carcinoma through nuclear factor-κB-mediated upregulation of vascular endothelial growth factor-C. Cancer Science, 105(10), 1261-1271.
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3

Western Blot Analysis of Cadherins

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Total protein (30 μg) was separated by 10% SDS-PAGE and transferred to PVDF membranes (Amersham Biosciences). The membranes were blocked in 1% bovine serum albumin and incubated with primary antibodies against R-cad at a dilution of 1:100 at 4°C for 10 h (Abnova, USA), β-actin at a dilution of 1:1000 at 4°C for 2 h (Santa Cruz, USA), and E-cad at a dilution of 1:1000 at 4°C for 10 h (Abcam, USA). After washing with TBST, alkaline phosphatase-conjugated secondary antibodies were added, and the protein bands were visualized using CDP-Star reagents (Roche, IN, USA).
Xie J., Feng Y., Lin T., Huang X.Y., Gan R.H., Zhao Y., Su B.H., Ding L.C., She L., Chen J., Lin L.S., Lin X., Zheng D.L, & Lu Y.G. (2016). CDH4 suppresses the progression of salivary adenoid cystic carcinoma via E-cadherin co-expression. Oncotarget, 7(50), 82961-82971.
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4

HBV Core DNA Detection and Analysis

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HBV core DNA was subjected to electrophoresis, transferred onto Hybond-XL membranes, and hybridized with digoxigenin-labelled probes generated using the PCR DIG Probe Synthesis Kit (11636090910, Roche, Switzerland). Signal detection was facilitated by Anti-Digoxigenin-Ap (11093274910, Roche, Switzerland), Fab fragments, and CDP-Star reagents (12041677001, Roche, Switzerland).
Wang J., Yuan X., Wang Y., Zhang Y., Han M., Lu H., Liu S., Zhang Y., Ge F., Liu Y, & Cheng J. (2024). PreS1BP mediates inhibition of Hepatitis B virus replication by promoting HBx protein degradation. Virus Research, 341, 199326.
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5

Immunoblotting Analysis of Protein Ubiquitination

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Cell lysates were prepared using RIPA lysis buffer (Pierce) containing a proteinase inhibitor cocktail (Roche Diagnostics). A total of 30 μg protein extracts were quantified and then subjected to electrophoresis on a 12% SDS-PAGE gel. The proteins were transferred to polyvinylidene difluoride (PVDF) membranes (Amersham Biosciences) and blocked in Tris-buffered saline (TBS) containing 2% bovine serum albumin (BSA). The specific antibodies used including anti-FLAG (Cell Signaling Technology), anti-USP15 (Santa Cruz), anti-K48-linkage Specific Polyubiquitin (Cell Signaling Technology), anti-ubiquitin (Cell Signaling Technology), anti-DDB1(Cell Signaling Technology), anti-USP5 (Santa Cruz), and anti-β-actin(Sigma Aldrich). Proteins were detected by addition of alkaline phosphatase (AP)-conjugated secondary antibody. Visualization of the immunoreactive proteins was performed by addition of CDP STAR reagents (Roche). Densitometry analysis of band signals was carried out using Image J software and the levels of HBx protein in cells transfected with pHBx-FLAG + pUSP15-myc were expressed as the relative intensity (RI) to that in the empty vector control pUSP5-myc or pHBx-FLAG after normalization to β-actin.
Su Z.J., Cao J.S., Wu Y.F., Chen W.N., Lin X., Wu Y.L, & Lin X. (2017). Deubiquitylation of hepatitis B virus X protein (HBx) by ubiquitin-specific peptidase 15 (USP15) increases HBx stability and its transactivation activity. Scientific Reports, 7, 40246.
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6

Quantitative Protein Expression Analysis

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Protein (30 μg) was subjected to 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and electrophoretically transferred to a polyvinylidene fluoride membrane (Millipore, Billerica, MA). Protein blots were incubated separately with antibodies against Dicer1 (No. 3363, 1:500, Cell Signaling Technology, Beverly, MA), FABP1 (HPA028275, 1:100, Sigma-Aldrich, St. Louis, MO), or β-actin (1:4000, Sigma-Aldrich), probed with the appropriate alkaline phosphatase-conjugated secondary antibody, and detected using chemiluminescence. Immunoreactive protein bands were visualized by adding CDP-Star reagents (Roche Diagnostics, GmbH). Band signal intensities were quantified using Quantity One densitometric software (Bio-Rad, Hercules, CA).
Wu Y.L., Zhu Y.B., Huang R.D., Peng X.E, & Lin X. (2017). Multiple MicroRNAs Ameliorate Hepatocyte Steatosis and Injury by Suppressing FABP1 Expression. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 44(6).
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7

Northern Blot Analysis of Transcripts

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Cell fractionation was performed as described previously (20 (link)). Total RNA was extracted (Nucleospin RNAII; Takara Bio) and poly(A)+ RNA was purified (NucleoTrap mRNA Mini kit; Takara Bio) according to the manufacturer's instructions. The RNA samples were loaded on 1.5% gels (NorthernMax-Gly Kit; Life Technologies), transferred to Hybond N+ nylon membranes (GE Healthcare) and probed with internally DIG-labeled sequences following pre-hybridization in ULTRAhyb hybridization buffer (Ambion). DIG-labeled RNA probes were prepared from template DNA amplified by specific primers (Supplementary Table S2) using a DIG Northern Starter Kit (Roche). Visualization of transcripts was performed with a CDP-Star reagent (Roche). Signals were detected by an LAS4000 mini biomolecular imager (GE Healthcare). The densitometric analysis of each band was performed by ImageQuant TL analysis software (GE Healthcare).
Koyama A., Sugai A., Kato T., Ishihara T., Shiga A., Toyoshima Y., Koyama M., Konno T., Hirokawa S., Yokoseki A., Nishizawa M., Kakita A., Takahashi H, & Onodera O. (2016). Increased cytoplasmic TARDBP mRNA in affected spinal motor neurons in ALS caused by abnormal autoregulation of TDP-43. Nucleic Acids Research, 44(12), 5820-5836.
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8

Western Blot Analysis of mEH Protein

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A total of 30 μg protein was subjected to 12% SDS-PAGE, and then electrophoretically transferred to a polyvinylidene fluoride (PVDF) membrane (Millipore, Billerica, MA, USA). Protein blots were incubated separately with a panel of specific antibodies such as anti-mEH (1:500 dilution, Santa Cruz Biotechnology) and anti-β-actin (1:4000 dilution, Sigma). An alkaline phosphatase (AP)-conjugated goat anti-mouse IgG was used as a secondary antibody. CDP-Star reagent (Roche Diagnostics) was used for color development.
Chen J.Y., Chen W.N., Jiao B.Y., Lin W.S., Wu Y.L., Liu L.L, & Lin X. (2014). Hepatitis B spliced protein (HBSP) promotes the carcinogenic effects of benzo [alpha] pyrene by interacting with microsomal epoxide hydrolase and enhancing its hydrolysis activity. BMC Cancer, 14, 282.
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9

RNA Extraction and Detection Protocol

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We extracted RNA from specific brain regions with Trizol reagent (Invitrogen) according to the manufacturer’s instructions. We loaded 5–10 μg total RNA into a urea-containing 12.5% acrylamide gel (Sequagel-Ureagel, National Diagnostics) and electrophoresed in standard TBE (1x) buffer. We then transferred RNA to a positively charged nylon membrane (Amersham) and hybridized overnight with digoxigenin-labeled LNA probes (Exiqon). We used anti-digoxigenin antibody linked to alkaline phosphatase (Roche, 11 093 274 910; 1:10,000) and CDP-star reagent (Roche) for detection.
Gascon E., Lynch K., Ruan H., Almeida S., Verheyden J., Seeley W.W., Dickson D.W., Petrucelli L., Sun D., Jiao J., Zhou H., Jakovcevski M., Akbarian S., Yao W.D, & Gao F.B. (2014). Alterations in microRNA-124 and AMPA receptors contribute to social behavioral deficits in frontotemporal dementia. Nature medicine, 20(12), 1444-1451.
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10

C9ORF72 Repeat Expansion Analysis by Southern Blot

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Southern blot analysis was used to analyze the C9ORF72 repeat expansion mutation as described69 (link). DNA was isolated using Wizard® genomic DNA purification kit (Promega). 20 µg of DNA was digested overnight with Xba endonuclease, separated on a 0.8% agarose gel electrophoresis, gel was subsequently depurinated, denatured, and transferred overnight on to a positively charged nylon membrane (Roche). The membrane was UV cross-linked and hybridized overnight at 47 °C with a digoxigenin (DIG)-labeled PCR probe. PCR probe was synthesized from genomic DNA using PCR DIG probe synthesis kit (Roche) using following primers South Fw: 5′-CTTGCAGATCAAAAGGCACA-3′ and South Rev: 5′-TTGACGCACCTCTCTTTCCT-3′. Following hybridization, membrane was washed and labeled with anti-DIG antibody and developed using CDP-Star reagent as per manufacturer’s recommendation (Roche).
Selvaraj B.T., Livesey M.R., Zhao C., Gregory J.M., James O.T., Cleary E.M., Chouhan A.K., Gane A.B., Perkins E.M., Dando O., Lillico S.G., Lee Y.B., Nishimura A.L., Poreci U., Thankamony S., Pray M., Vasistha N.A., Magnani D., Borooah S., Burr K., Story D., McCampbell A., Shaw C.E., Kind P.C., Aitman T.J., Whitelaw C.B., Wilmut I., Smith C., Miles G.B., Hardingham G.E., Wyllie D.J, & Chandran S. (2018). C9ORF72 repeat expansion causes vulnerability of motor neurons to Ca2+-permeable AMPA receptor-mediated excitotoxicity. Nature Communications, 9, 347.
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