Abcam
IGF1 Receptor Human ELISA Kit
ab100546
Info: View Product View Product Specs
Our lab deals with molecular imaging of cancer, but we must first know the expression of a specific target to choose the correct tumor-derived animal models. For this reason, we perform Western blots or ELISAs. In this study, we needed to determine the expression levels of IGF1R in three pancreatic cancer cell lines, known as MiaPaCa-2, BxPC-3, and AsPC-1. We chose this product because Abcam has always been a great company to us. There are several IGF1R ELISA kits on the market.
Determination of IGF-1R expression by ELISA
Cell lysate
The IGF-1R human ELISA kit (Abcam, Cambridge, Massachusetts, USA) was used to quantify the amount of human IGF-1R in three pancreatic cancer cell lines. The ELISA was performed according to the manufacture’s protocol. Briefly, cells were first lysed with RIPA buffer and total protein was measured as described for the Western blot. IGF-1R standards were made from 0 to 6000 pg/mL. Next, 100 µL of standards and samples were added into appropriate wells. The plate was covered and incubated for 12 h at 4 ºC with gentle shaking. The solution was discarded and wells were washed four times with 1X Wash Solution. Next, 100 µL of 1X Biotinylated IGF1R Detection Antibody was added to each well. The plate was incubated for 1 h with gentle shaking at room temperature. The solution was removed and the plate was washed four times with 1X Wash Solution. Next, 100 µL of 1X HRP-Streptavidin solution was added to each well and the plate was incubated for 45 min at room temperature. The solution was discarded and the plate was washed four times with 1X Wash Solution. Next, 100 µL of TMP One Step Substrate Reagent was added to each well and the plate was incubated in the dark for 30 min at room temperature. Lastly, 50 µL of Stop Solution was added to the corresponding wells and measurements were taken at 450 nm.
Don't reuse pipette tips as you load your samples. Good pipetting techniques are essential for good results.
After making a standard curve, we determined the concentration of IGF1R in our samples. The calculated values were 9.6 +/- 0.15, 15.9 +/- 1.2, and 3.36 +/- 0.27 picograms per 25 micrograms of total protein for MiaPaCa-2, BxPC-3, and AsPC-1, respectively.
None
Simple to use with great instructions
Take a long time
I highly recommend this ELISA kit from Abcam. In the future, I will continue to use ELISA kits from this company.
Product Name: IGF1 Receptor Human ELISA KitSupplier Page