Immunohistochemistry Estrogen receptor (ER) Rabbit

Get tips on using Cleaved Notch1 (Val1744) (D3B8) Rabbit mAb #4147 to perform Immunohistochemistry Mouse - Notch1

Get tips on using Lab Vision™ Ki-67, Rabbit Monoclonal Antibody to perform Immunohistochemistry Mouse - Ki67

Get tips on using NF-κB p65 (D14E12) XP® Rabbit mAb #8242 to perform Immunohistochemistry Mouse - NFκB / p65

Get tips on using OSTEOPONTIN (O-17) ANTI-HUMAN RABBIT IGG AFFINITY PURIFY to perform Immunohistochemistry Mouse - Spp1/OPN

Get tips on using Anti Type X Collagen (raised against rat) pAb (Rabbit, Antiserum) to perform Immunohistochemistry Mouse - Col X

Get tips on using GR Antibody (G-5): sc-393232 to perform Immunohistochemistry Human - GR/glucocorticoid receptor

Get tips on using Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb #8814 to perform Immunohistochemistry Mouse - β-Catenin

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

miRNA is the inherent gene silencing machinery which can have more than one mRNA target, whereas siRNA can be designed to target a particular mRNA target. By design, both siRNA and miRNA are 20-25 nucleotides in length. The target sequence for siRNAs is usually located within the open reading frame, between 50 and 100 nucleotides downstream of the start codon. There are two ways in which cells can be transfected with desired RNAi: 1. Direct transfection (with calcium phosphate co-precipitation or cationic lipid mediated transfection using lipofectamine or oligofectamine), and 2. Making RNAi lentiviral constructs (followed by transformation and transduction). Lentiviral constructs are time consuming, but provide a more permanent expression of RNAi in the cells, and consistent gene silencing. Direct transfection of oligonucleotides provides temporary genetic suppression. Traditional methods like calcium phosphate co-precipitation have challenges like low efficiency, poor reproducibility and cell toxicity. Whereas, cationic lipid-based transfection reagents are able to overcome these challenges, along with applicability to a large variety of eukaryotic cell lines. When using oligos, the ideal concentration lies between 10-50nM for effective transfection.