The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 0.1 - 0.2 µg/ml. To detect hEGF by Western Blot analysis this antibody can be used at a concentration of 0.1 - 0.2 µg/ml. Used in conjunction with compatible secondary reagents the detection limit for recombinant hEGF is 1.5 - 3.0 ng/lane, under either reducing or non-reducing conditions. Western blot bands should be around 6.2 kDa.
Use a concentration of 0.5 µg/ml. To detect hEGF by direct ELISA (using 100µl/well antibody solution) a concentration of at least 0.5µg/ml of this antibody is required. This antigen affinity purified antibody, in conjunction with compatible secondary reagents, allows the detection of 0.2 - 0.4 ng/well of recombinant hEGF.
Use a concentration of 0.25 - 0.35 µg/ml. To yield one-half maximal inhibition [ND50] of the biological activity of hEGF (2.0 ng/ml), a concentration of 0.25 - 0.35 µg/ml of this antibody is required.
Use a concentration of 0.5 µg/ml.
For sandwich ELISA, use this antibody as Detection at 0.5µg/ml with ab18635 as Capture.
Use a concentration of 1 µg/ml.
EGF stimulates the growth of various epidermal and epithelial tissues in vivo and in vitro and of some fibroblasts in cell culture. Magnesiotropic hormone that stimulates magnesium reabsorption in the renal distal convoluted tubule via engagement of EGFR and activation of the magnesium channel TRPM6. Can induce neurite outgrowth in motoneurons of the pond snail Lymnaea stagnalis in vitro (PubMed:10964941).
Expressed in kidney, salivary gland, cerebrum and prostate.
Contains 9 EGF-like domains. Contains 9 LDL-receptor class B repeats.
O-glycosylated with core 1-like and core 2-like glycans. It is uncertain if Ser-954 or Thr-955 is O-glycosylated. The modification here shows glycan heterogeneity: HexHexNAc (major) and Hex2HexNAc2 (minor).
ab9695 at 1.0 µg/ml staining normal human skin by IHC-P. The antibody was incubated overnight at 4˚C. An HRP-labeled polymer detection system was used with a non-alcohol soluble AEC chromogen and a proteinase K antigen retrieval.
Western blot - Anti-EGF antibody (ab9695)
Primary ab9695 was used at a dilution of 0.2 ug/ml.
Lane 1: 250 ng Recombinant human EGF protein
Lane 2: 125 ng Recombinant human EGF protein
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGF antibody (ab9695)This image is courtesy of an anonymous Abreview
Immunohistochemical analysis of murine forepaw vessel, staining EGF with ab9695.
Tissue was fixed with formaldehyde and blocked with 1% blocking solution for 15 minutes at room temperature; antigen retrieval was by proteinase K. Samples were incubated with primary antibody (4 µg/ml in BSA in TBS) for 1 hour. An undiluted HRP-conjugated goat anti-rabbit polyclonal IgG was used as the secondary antibody.
Zhang B et al. Metastasis-associated protein 2 promotes the metastasis of non-small cell lung carcinoma by regulating the ERK/AKT and VEGF signaling pathways. Mol Med Rep17:4899-4908 (2018).
Read more (PubMed: 29393472) »
Fang C et al. Berberine ameliorates neonatal necrotizing enterocolitis by activating the phosphoinositide 3-kinase/protein kinase B signaling pathway. Exp Ther Med15:3530-3536 (2018).
Read more (PubMed: 29545879) »