Sheep anti digoxigenin ap

Manufactured by Roche

Sheep anti-Digoxigenin-AP is a laboratory reagent used for the detection and quantification of digoxigenin-labeled molecules in various biological assays. It is an antibody conjugated with alkaline phosphatase, which can be used to generate a colorimetric or chemiluminescent signal when bound to digoxigenin-labeled targets.

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Lab products found in correlation

Lna microrna ish mir 122 optimization kit, by Qiagen (2 mentions) Nuclear fast red, by Vector Laboratories (2 mentions) Nbt bcip, by Vector Laboratories (2 mentions) Nbp2 14833, by Novus Biologicals (2 mentions) Fast red, by Merck Group (1 mentions) Tyramide alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Pcr xl topo, by Thermo Fisher Scientific (1 mentions) Tyramide signal amplification kit tsa, by Thermo Fisher Scientific (1 mentions) Lsm 700 axioimager, by Zeiss (1 mentions) Goat anti mouse hrp, by Thermo Fisher Scientific (1 mentions) Nbt bcip stock solution, by Roche (1 mentions) Dig rna labeling kit, by Roche (1 mentions) Rabbit anti β gal, by Thermo Fisher Scientific (1 mentions) Mouse anti brdu g3g4, by Developmental Studies Hybridoma Bank (1 mentions)

5 protocols using sheep anti digoxigenin ap

Dual-Label In Situ Hybridization for vg and ncRNA

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PCR products from embryonic cDNA for the vg gene or the vg PRE/TRE ncRNA were cloned in both orientations into PCR XL Topo (Invitrogen). For in situ hybridization, RNA probes were in vitro transcribed from each cDNA strand. One probe was Dig labeled (Boehringer), detected with sheep Anti-Digoxigenin-AP (Roche) and visualized with FastRed (Sigma). The other probe was labeled with fluorescein and detected with primary antibody: Mouse anti-fluorescein (Roche), secondary antibody: goat anti-mouse-HRP (Invitrogen) and visualized with Alexa Fluor 488 Tyramide (Invitrogen) using Tyramide Signal Amplification kit (TSA™, Invitrogen) according to manufacturers instructions. Label swaps were performed to ensure specificity. In the images shown, vg mRNA was detected with Fast Red, and PRE/TRE transcripts were detected with TSA. Images were taken using confocal microscopy, with LSM 700 Axioimager (larval tissues) or LSM 510 Axiovert 200M (embryos) (Zeiss).

Herzog V.A., Lempradl A., Trupke J., Okulski H., Altmutter C., Ruge F., Boidol B., Kubicek S., Schmauss G., Aumayr K., Ruf M., Pospisilik A., Dimond A., Senergin H.B., Vargas M.L., Simon J.A, & Ringrose L. (2014). A strand-specific switch in noncoding transcription switches the function of a Polycomb/Trithorax response element. Nature genetics, 46(9), 973-981.

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In Situ Hybridization of Fst, Ran, and Ube2c

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After linearization of plasmids bearing hamster Fst, Ran and Ube2c cDNAs, digoxigenin-labeled antisense and sense cRNA probes were transcribed in vitro using the DIG RNA Labeling Kit (Roche Applied Science, Mannheim, Germany). In situ hybridization with digoxigenin-labeled probe was performed as described previously (Lei et al. 2012 (link)). After fixing with 4% paraformaldehyde solution in PBS, frozen sections were hybridized with antisense or sense cRNA probes at 55°C for 16 h, respectively. Sections were then washed and incubated in sheep anti-Digoxigenin-AP (1:5,000; Roche Applied Science). The signal was visualized with diluted NBT/BCIP Stock Solution (Roche Applied Science). All of the sections were counterstained with 1% methyl green.

Lei W., Herington J., Galindo C.L., Ding T., Brown N., Reese J, & Paria B.C. (2014). Cross-species transcriptomic approach reveals genes relating to hamster implantation sites. Reproduction (Cambridge, England), 148(6), 607-621.

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Immunostaining of Drosophila Wing Discs

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Third instar larvae were dissected in cold phosphate-buffered saline (PBS) and fixed in 4% formaldehyde/PBS for 20 min at room temperature. They were then washed and permeabilized in PBT (0.2% Triton X-100 in PBS) for 30 min and blocked in BBT (0.3% BSA, 250 mM NaCl in PBT) for 1 h. Samples were incubated overnight at 4°C with primary antibody diluted in BBT, washed three times (15 min each) in BBT and incubated with secondary antibodies and DAPI (1 μg/ml) for 1.5 hour at room temperature. After three washes with PBT (15 min each), wing discs were placed in mounting medium (80% glycerol/PBS containing 0.05% n-Propyl-Gallate). All steps were performed on a rocking platform at the indicated temperature. The following primary antibodies were used: mouse anti-BrdU (G3G4; Developmental Studies Hybridoma Bank (DSHB)); mouse anti-MMP1 (3A6B4, DSHB); mouse anti-p53 (7A4, DSHB); rabbit anti-p-Histone H3 (sc-8656, Santa Cruz); rabbit anti-β-Gal (A11132, Invitrogen); and sheep anti-Digoxigenin-AP (#11093274910, Roche). The following secondary antibodies were used: anti-mouse IgG-Alexa Fluor 594; anti-mouse IgG-Alexa Fluor 488; anti-rabbit IgG-Alexa Fluor 594; and anti-rabbit IgG-Alexa Fluor 488 (Jackson InmunoResearch).

Sanchez J.A., Mesquita D., Ingaramo M.C., Ariel F., Milán M, & Dekanty A. (2019). Eiger/TNFα-mediated Dilp8 and ROS production coordinate intra-organ growth in Drosophila. PLoS Genetics, 15(8), e1008133.

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Immunohistochemistry and In Situ Hybridization Protocols

Cited 1 time

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IHC was performed as previously described^{69 (link)} using a 1:400 antibody dilution for PKM2 (C-11), 1:600 dilution for PKM1 (Cat# NBP2-14833, Novus Biologicals; Littleton, CO), and 1:250 dilution for GLUT1 (AbCam; Cat# ab652). ISH was performed as described^{70 (link)} using LNA^™ microRNA ISH miR-122 optimization kit (Cat# 90003, Exiqon; Woburn, MA) followed by incubation of sheep anti-digoxigenin-AP (Cat# 11093274910, Roche Diagnostics; Mannheim, Germany), and developed with NBT:BCIP (Cat# SK-5400, Vector Laboratories; Burlingame, CA) at 30°C overnight. Nuclear fast red was used to counterstain nuclei (Cat# H-3403, Vector Laboratories).

Fong M.Y., Zhou W., Liu L., Alontaga A.Y., Chandra M., Ashby J., Chow A., O’Connor S.T., Li S., Chin A.R., Somlo G., Palomares M., Li Z., Tremblay J.R., Tsuyada A., Sun G., Reid M.A., Wu X., Swiderski P., Ren X., Shi Y., Kong M., Zhong W., Chen Y, & Wang S.E. (2015). Breast cancer-secreted miR-122 reprograms glucose metabolism in pre-metastatic niche to promote metastasis. Nature cell biology, 17(2), 183-194.

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Immunohistochemistry and In Situ Hybridization Protocols

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