For detection of Lgr5 and Olfm4 mRNA, we designed riboprobes targeting the coding sequence to 3’UTR region of these genes (Supplemental Table 5 ). Targeted probe region DNA were amplified using standard PCR, with the amplicon length of about 500 bp. These DNA amplicons were used to generate DIG-labeled riboprobes using T7 RNA polymerase. For staining, Formalin fixed paraffin embedded tissue sections were deparaffinized with Citrisolv, washed in 100% ethanol, and air dried. Slides were blocked with Boeringer Blocking Reagent (Roche) and yeast tRNA at 68°C for 1 hour. 0.5 ng/mL probes were hybridized to sections at 68°C overnight. After washing, sections were again blocked with Boeringer Blocking Reagent and 10% sheep serum at room temperature for 1 hour. Alkaline phosphatase labeled sheep anti-DIG antibody (Roche) was incubated with sections at 1:2000 dilution at 4°C overnight. After washing, positive signals were developed using NBT/BCIP substrate (Thermo). Slides were then fixed in 10% neutral buffered formalin and mounted with aqueous mounting medium.
Alkaline phosphatase labeled sheep anti dig antibody
Manufactured by Roche
Alkaline phosphatase labeled sheep anti-DIG antibody is a laboratory reagent used to detect and quantify the presence of digoxigenin (DIG) in samples. It consists of an antibody produced in sheep that binds to DIG, with an alkaline phosphatase enzyme attached. This enzyme can catalyze a colorimetric reaction, allowing for the visualization and measurement of DIG-labeled targets.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using alkaline phosphatase labeled sheep anti dig antibody
1
In Situ Hybridization of Lgr5 and Olfm4
2
In Situ Hybridization of Lgr5 and Olfm4
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For detection of Lgr5 and Olfm4 mRNA, we designed riboprobes targeting the coding sequence to 3’UTR region of these genes (Supplemental Table 5 ). Targeted probe region DNA were amplified using standard PCR, with the amplicon length of about 500 bp. These DNA amplicons were used to generate DIG-labeled riboprobes using T7 RNA polymerase. For staining, Formalin fixed paraffin embedded tissue sections were deparaffinized with Citrisolv, washed in 100% ethanol, and air dried. Slides were blocked with Boeringer Blocking Reagent (Roche) and yeast tRNA at 68°C for 1 hour. 0.5 ng/mL probes were hybridized to sections at 68°C overnight. After washing, sections were again blocked with Boeringer Blocking Reagent and 10% sheep serum at room temperature for 1 hour. Alkaline phosphatase labeled sheep anti-DIG antibody (Roche) was incubated with sections at 1:2000 dilution at 4°C overnight. After washing, positive signals were developed using NBT/BCIP substrate (Thermo). Slides were then fixed in 10% neutral buffered formalin and mounted with aqueous mounting medium.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!