Immunoblotting and dot blotting
. J Immunol Methods.
The biotin binding activities of the wild type and the mutant streptavidins were evaluated by dot blotting
using alkaline phosphatase-labeled biotin.
IgE-reactivity of forty seven serum samples from allergic Puerto Rican patients was analyzed by dot blotting
assays using recombinant allergens from mites.
With regard to the regulation of survival pathway, using dot blotting, we found cardiac insulin-like growth factor-1 (IGF-1) and IGF-1 receptor mRNA levels to be significantly increased, indicating that compensative effects of IGF-1 survival signaling could occur.
To understand whether the effects of SHS on rat hearts are mediated through activating apoptotic pathways, including mitochondria-dependent and Fas death-receptor-dependent signalings, or through suppressing survival pathways, we examined the cardiac levels of signaling proteins and gene expression in these pathways by performing reverse-transcriptase polymerase chain reaction (RT-PCR), Western blotting, or dot blotting. We used these results to explore the molecular mechanisms of the pathogenesis of cardiac disease induced by cigarette smoke.
RNA dot blotting. We used RNA dot blotting for the hybridization and detection of IGF-1 and IGF-1R mRNAs as described previously (Huang et al.
Western and dot blotting were conducted using standard methods and controls .
The protein that was eluted following removal of [Ca.sup.+2] was identified as coagulogen by molecular mass ([sim]21 kDa) and reactivity with anti-coagulin antibodies (Western blotting and dot blotting).
The supernatant fraction contained substantial quantities of [[alpha].sub.2]M, as shown by Western blotting and dot blotting using an affinity-purified antibody to Limulus [[alpha].sub.2]M.
Dot blotting requires positive-control samples to monitor the sensitivity and specificity of each diagnostic blot.
The integrity of this product as positive-control DNA for the purposes of dot blotting was validated by probing a PCR product from a patient known to carry the A4136G base substitution in parallel dot blotting experiments.
For dot blotting
sap extractions were prepared by grinding 50 mg of fresh plant tissues using the previous protocol for preparation of DNA as described by Arismendi et al.2010.