siRNA / miRNA gene silencing Human PC3 (human prostate cancer cell line)

Get tips on using Human Dkk-1 ELISA Kit (RAB0143) to perform ELISA Human - Dkk-1

Get tips on using Human Dkk-1 DuoSet ELISA (DY1906) to perform ELISA Human - Dkk-1

Get tips on using Human Cytochrome c Quantikine ELISA Kit to perform ELISA Human - Cytochrome C

Get tips on using Human ANGPTL3 (highly sensitive) Assay Kit (27750 ) to perform ELISA Human - Angiopoietin-Like 3 (AngptL3)

Get tips on using SurePrint G3 Human CGH Microarray Kit, 4x180K to perform Microarray Comperative genomic hybridization - Human MDA-MB-361

Get tips on using SurePrint G3 Human CGH Microarray Kit, 4x180K to perform Microarray Comperative genomic hybridization - Human MDA-MB-453

Get tips on using Human ICAM1 ELISA Kit (CD54) (ab100640) to perform ELISA Human - ICAM-1/CD54

Protein isolation is a technique that involves isolation and/ or purification of protein from cells or tissues via chromatography or electrophoresis. The major challenges in protein isolation include: 1. The concentration of proteins in cells is variable and tends to be small for some intracellular proteins. Unlike nucleic acids, proteins cannot be amplified. 2. Proteins are more unstable than nucleic acids. They are easily denatured under suboptimal temperature, pH or salt concentrations. 3. Finally, no generalized technique/protocol can be applied for protein isolation. Proteins may have different electrostatic (number of positively or negatively charged amino acids) or hydrophobic properties. Therefore, protein purification requires multiple steps depending on their charge (a negatively charged resin/column for positively charged proteins and vice-versa), dissolution (using detergents) and unlike in the case of DNA and RNA, instead of using salts, proteins should be isolated by isoelectric precipitation.

ELISA is the most commonly used method of detecting and quantifying the concentration of an antigen in an unknown sample. During the experiment, If you get a weak signal, then make sure reagents are at room temperature before starting the assay. Try increasing incubation times to ensure maximal antibody binding and amplify the signal. Secondly, if you get values above 0 in the negative control indicates a high background signal. Try to consider reducing your antibody concentration and prevent non-specific binding of antibodies by using affinity-purified antibody and suitable blocking buffers. To avoid high well to well variation, do not stack plates during incubation, no bubbles in the plate and wash wells thoroughly to avoid variation.

Get tips on using PE anti-human CD96 (TACTILE) Antibody to perform Flow cytometry Anti-bodies Human - CD96