Recombinant人EGF protein (Animal Free) (ab9697)

概述

描述

  • 性质Recombinant
  • 来源Escherichia coli
  • 氨基酸序列
    • AccessionQ6QBS2
    • 种属Human
    • 序列NSDSECPLSHDGYCLHDGVCMYIEALDKYACNCVVGYIGERCQYRDLKWW ELR
    • 分子量6 kDa
    • 氨基酸1 to 53
    • 额外的序列信息ab9697 is a globular protein containing 53 amino acid residues, including 3 intramolecular disulfide bonds.

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技术指标

Our Abpromise guarantee covers the use of ab9697 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

  • 生物活性

    The ED50 was determined by a cell proliferation assay using balb/c 3T3 cells is ≤ 0.1 ng/ml, corresponding to a specific activity of ≥ 1 x 107 units/mg.

  • 应用

    Western blot

    Cellular Activation

    SDS-PAGE

  • 内毒素水平< 0.010 Eu/µg
  • 形式Lyophilised
  • 补充说明Manufactured using all non-animal reagents.
  • Concentration information loading...

制备和贮存

  • 稳定性和存储

    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.

    n/a

    This product is an active protein and may elicit a biological response in vivo, handle with caution.

  • 复溶Centrifuge the vial prior to opening. Reconstitute in 500ul sterile filtered water to a concentration of 1mg/ml.

常规信息

  • 别名
    • Beta urogastrone
    • EGF
    • EGF_HUMAN
    • Epidermal growth factor
    • HOMG4
    • OTTHUMP00000219721
    • OTTHUMP00000219722
    • Pro epidermal growth factor
    • URG
    • Urogastrone
    see all
  • 功能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).
  • 组织特异性Expressed in kidney, salivary gland, cerebrum and prostate.
  • 疾病相关Hypomagnesemia 4
  • 序列相似性Contains 9 EGF-like domains.
    Contains 9 LDL-receptor class B repeats.
  • 翻译后修饰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).
  • 细胞定位Membrane.
  • Information by UniProt

Recombinant human EGF protein (Animal Free) 图像

  • Lane 1 :
    Lane 2 :
    Lane 3 :

    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
    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

    This image is courtesy of an anonymous Abreview

    Blocking Step: 5% milk for 1 hour at 25°C

    See Abreview

  • 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.

Recombinant human EGF protein (Animal Free) (ab9697)参考文献

This product has been referenced in:

See all 2 Publications for this product

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The two active proteins are essentially the same, and both can be used to induce phosphorylation as they have been tested in functional studies. The only difference would be that ab9697 is supplied as lyophilized powde...

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Application Western blot

Review text: Human skin fibroblasts (HFF-1) were starved in serum-free medium overnight, and then were incubated 30 min in the presence of either 25 or 50 ng/ml active EGF protein, doses widely used in stem cell culture medium, to test whether EGFR downstream signaling pathway is able to be activated.
Sample: Human Cell lysate - whole cell (skin fibroblasts)
Loading amount: 20 µg
Specification: skin fibroblasts
Treatment: 25 and 50 ng/ml EGF for 30 min
Gel Running Conditions: Non-reduced Denaturing (10%)
Blocking step: Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Other product details
Dilution: 1/1000
Incubation time: 16 hour(s) and 0 minute(s) · Temperature: 4°C

Primary antibody (in addition to 'EGF protein (Active)')
Name: Non-Abcam antibody was used: phospho-p44/42 MAPK (Erk1/2)
Host species: Rabbit
Clonality: Polyclonal
Dilution: 1/1000

Secondary antibody
Name: Non-Abcam antibody was used: anti-rabbit 2nd Ab HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution: 1/10000

Detection
Detection method: ECL
Exposure: 1 second(s)
Bands: Specific: 42/44 kDa

Additional data
Additional Notes: Incubation of human fibroblasts with active EGF protein for short time greatly activated EGFR signaling pathway, as the receptor underwent endocytosis and dramatic phosphorylation of p44/42 MAPK.
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提交于 Jun 25 2010

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