rna-isolation-purification-cells-primary-mouse-ventricles

Get tips on using CD11b Antibody, anti-human, PE, REAfinity™ to perform Flow cytometry Anti-bodies Human - CD11b

Get tips on using RealTime-Glo™ Annexin V Apoptosis and Necrosis Assay to perform Necrosis HCT 116

Get tips on using GenLadder 50bp (ready-to-use) with dye Orange G to perform DNA Ladder 50 bp

Get tips on using CD326 (EpCAM) Antibody, anti-human, PE, REAfinity™ to perform Flow cytometry Anti-bodies Human - CD326/EpCAM

Get tips on using GenLadder 100 bp + 1.5 kbp (ready-to-use), DNA marker to perform DNA Ladder 100 bp

Get tips on using CD163 Antibody, anti-human, PE-Vio® 770, REAfinity™ to perform Flow cytometry Anti-bodies Human - CD163

Get tips on using CD171 (L1CAM) Antibody, anti-human, PE-Vio® 770, REAfinity™ to perform Flow cytometry Anti-bodies Human - CD171/L1CAM

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.

In ChIP, the most vital step is the binding of an antibody and choosing the right antibody. The binding affinity of different types of immunoglobulins to protein A or G differs significantly. Henceforth, it is recommended to choose either protein A or protein G coated beads. If you do not see any product in the positive control, add 5–10 μg of chromatin and 1–5 μg of antibody to each IP reaction and incubate with antibody overnight and an additional 2 hr after adding Protein G/A beads. If no product in the experimental, add more DNA to the PCR reaction or increase the number of amplification cycles. Choose an alternate, ChIP-validated antibody if the antibody does not work.

In ChIP, the most vital step is the binding of an antibody and choosing the right antibody. The binding affinity of different types of immunoglobulins to protein A or G differs significantly. Henceforth, it is recommended to choose either protein A or protein G coated beads. If you do not see any product in the positive control, add 5–10 μg of chromatin and 1–5 μg of antibody to each IP reaction and incubate with antibody overnight and an additional 2 hr after adding Protein G/A beads. If no product in the experimental, add more DNA to the PCR reaction or increase the number of amplification cycles. Choose an alternate, ChIP-validated antibody if the antibody does not work.