EGF Antibody (3A8)

EGF Monoclonal Antibody for ELISA, IF, Western blot

>> See 12 other antibodies for EGF (HMGF (human milk growth factor), PGF (prostatic growth factor), tooth-lid factor, Urogastrone)
Synonyms:
HMGF (human milk growth factor), PGF (prostatic growth factor), tooth-lid factor, Urogastrone
Entrez Gene ID:
(Hu) 1950
UniProt ID:
(Hu) P01133
Details
Host / Isotype: Mouse / IgG
Class: Monoclonal
Type: Antibody
Clone: 3A8
Species Reactivity: Human (Hu)
Immunogen: Recombinant Human EGF
Ordering Information
Pierce EGF Antibody (3A8)
Product #
M806
Size
200 µg
Price
$299.00
Purchase
Add EGF Antibody (3A8) to your cart.
Storage: -20°C
Form: 200 µg of protein A purified antibody (1 mg/ml) in PBS, preservative- and carrier-free.


Applications Dilution *
Western Blot (WB) 1:1000 - 1:5000
Immunofluorescence (IF) 1:50
ELISA (ELISA) 0.125 - 0.25 µg/ml
* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
Product Specific Information
The M806 anti-EGF antibody (Clone 3A8) has successfully been paired as the detection antibody in a sandwich ELISA with coating antibody M805 (Clone 1H11).

Typical dilutions for sandwich ELISA include 1-3 µg/ml for coating and 0.125-0.5 µg/ml for detection.
General Information
The protein encoded by EGF gene is a growth factor that stimulates cell growth, proliferation, and differentiation by binding to its receptor EGFR. Human EGF is a 6045-Da protein with 53 amino acid residues and three intramolecular disulfide bonds
Product Images
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  • EGF Antibody (M806) in ELISA
    ELISA with anti-EGF Monoclonal Antibody [3A8] (M806)

    ELISA with anti-EGF Monoclonal Antibody [3A8] (M806)

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    Sandwich ELISA analysis of Human EGF was performed using the Thermo Scientific ELISA Kit (Product # EHEGF) by coating a blank 96-well microtiter plate with 100µl per well of Human EGF moncolonal antibody (Product # M805) in duplicate at 1, 3, 4, 5, 7, and 9µg/ml in DPBS (Product # 28374) and incubating for 12-18 hours at 4C. The plate was aspirated and blocked with 300µl per well of 4% BSA and 5% sucrose in DPBS for 1 hour at room temperature. Human EGF recombinant protein at 50µl per well was added in duplicate at 500, 250, 125, 62.5, 31.25, 15.625, 7.8125 and 0 pg/ml for 2 hours at room temperature along with 50µl of Human EGF biotinylated monoclonal antibody (Product # M806B) in all applicable wells at 0.2µg/ml for 2 hours at room temperature. The plate was washed with ELISA Wash Buffer (Product # N503) and incubated with 100µl per well of Streptavidin-HRP (Product # N504) in all test wells at 1:130,000 dilution for 1 hour at room temperature and then washed and incubated with 100µl per well of TMB substrate (Product # 34028) for 30 minutes at room temperature in the dark. The plate was stopped with 0.16M sulfuric acid (Product # N600). Absorbances were read on a spectrophotometer at 450-550nm
     
  • EGF Antibody (M806) in IF
    Immunofluorescence with anti-EGF Monoclonal Antibody [3A8] (M806)

    Immunofluorescence with anti-EGF Monoclonal Antibody [3A8] (M806)

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    Immunofluorescent analysis of EGF (green) in HeLa cells either left untreated (left panel) or treated with 10ng/ml EGF (right panel) for 5 minutes. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with an EGF monoclonal antibody (Product # M806) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
     
  • EGF Antibody (M806) in WB
    Western Blot with anti-EGF Monoclonal Antibody [3A8] (M806)

    Western Blot with anti-EGF Monoclonal Antibody [3A8] (M806)

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    Western blot analysis of Human EGF was performed by loading 2ug of recombinant human EGF (Lane 1) or supernatant from an EGF expression clone transfected in 293T cells onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with an EGF monoclonal antibody recognizing Human EGF (Product # M806) at a dilution of 1:5000 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a goat anti-rabbit-HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for at least one hour. Membranes were washed and chemiluminescent detection performed using Super Signal West Dura (Product # 34075).
     
 
 
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