Immunohistochemistry Alpha smooth muscle Actin Rabbit Mouse

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 Mouse CXCL12/SDF-1 alpha Quantikine ELISA Kit to perform ELISA Mouse - SDF-1/CXCL12

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary rabbit aortic smooth muscle cells

Get tips on using Estrogen Receptor alpha Antibody (F-10): sc-8002 to perform Immunohistochemistry Mouse - ERα

Get tips on using β-Actin Antibody #4967 to perform Western blotting β-Actin

Get tips on using Anti-beta Actin antibody (ab8227) to perform Western blotting β-Actin

Isolating RNA from tissues and paraffin-embedded tissue samples can be challenging due to cross-linking of biomolecules and fragmented nucleic acids. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in presence of RNAse inhibitors. The homogenization process should be carried out on dry ice to maintain the integrity of RNA.

Get tips on using Recombinant Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063) to perform Immunohistochemistry Mouse - ERα

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.

Get tips on using PDGF Receptor β (28E1) Rabbit mAb to perform Immunohistochemistry PDGFβR - Rabbit Mouse -NA-