Recombinant human EGF protein (Animal Free) (ab9697)
Key features and details
- Expression system: Escherichia coli
- Purity: >= 98% SDS-PAGE
- Endotoxin level:
- Active: Yes
- Suitable for: WB, Cellular Activation, SDS-PAGE, HPLC
Preparation and Storage
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Stability and Storage
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
This product is an active protein and may elicit a biological response in vivo, handle with caution.
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ReconstitutionCentrifuge the vial prior to opening. Reconstitute in 500ul sterile filtered water to a concentration of 1mg/ml.
Images
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Western blot - Recombinant human EGF protein (Animal Free) (ab9697) This image is courtesy of an anonymous AbreviewAll lanes :
Lane 1 : Whole cell lysate of human skin fibroblasts starved overnight in serum-free medium
Lane 2 : Whole cell lysate of human skin fibroblasts starved overnight in serum-free medium and then incubated for 30 min with 25 ng/ml active EGF
Lane 3 : Whole cell lysate of human skin fibroblasts starved overnight in serum-free medium and then incubated for 30 min with 50 ng/ml active EGF
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : An HRP-conjugatede Goat anti-rabbit IgG polyclonal at 1/10000 dilution
Developed using the ECL technique.
Performed under non-reducing conditions.
Observed band size: 42,44 kDa why is the actual band size different from the predicted?
Exposure time: 1 second
Blocking Step: 5% milk for 1 hour at 25°C
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Immunocytochemistry/ Immunofluorescence - Recombinant human EGF protein (Animal Free) (ab9697)Immunocytochemistry/ Immunofluorescence analysis of A431 (Human epidermoid carcinoma cell line) cells labeling EGFR with ab9697 at 1/100, 3 μg/ml. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077, a AlexaFluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1000, 2 μg/ml. Cells were counterstained with ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200, 2.5 μg/ml. Nuclear stain was DAPI (blue).
The green staining on the membrane was increased in the EGF (100ng/ml, 10min) treated A431 cells when compared with A431 cells without treatment. After LP treatment, the green signaling was obviously decreased.
For the pan antibody, there was no great difference after EGF (100ng/ml, 10min) or EGF (100ng/ml, 10min) + LP treatment. The data showed mostly membranous staining.
