discontinued

BM2207 EGF antibody

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Mouse anti Mouse EGF E5

BM2207

Product Description for EGF

Mouse anti Mouse EGF E5.
Presentation: Supernatant
Product is tested for Frozen Sections, Enzyme Immunoassay, Paraffin Sections.

Properties for EGF

Product Category Primary Antibodies
Quantity 1 ml
Synonyms Beta urogastrone antibody, Epidermal Growth Factor antibody, Pro epidermal growth factor antibody, URG antibody, Urogastrone antibody
Presentation Supernatant
Reactivity Ms
Applications C, E, P
Clonality Monoclonal
Clone E5
Host Mouse
Isotype IgG1
Shipping to Worldwide
PDF datasheet View Datasheet
Manufacturer Acris Antibodies GmbH
Material safety datasheet MSDS for Monoclonal Antibodies (de)

Datasheet Extract

Application ELISA (10 ng EGF is detectable) and spot blots (1 ng is detectable).
Use in formalin fixed frozen sections (1:20) and paraffin sections of salivary glands. See Protocol for more details.
Concentration 20 µg/ml
Protocols Indirect Immunoperoxidase Staining On Frozen Sections :
1. 4 to 6 micron thick sections should be used.
2. Sections are thawed, 1-2 hours at room temperature.
3. Tissue is fixed in acetone, 10 minutes.
4. Wash with PBS, 2 x 3 minutes.
5. Incubate with monoclonal antibody (diluted in PBS), 1-2 hours at room temperature.
6. Wash with PBS, 3 x 3 minutes.
7. Incubate with peroxidase labeled second antibody, 30-60 minutes at room temperature.
8. Wash with PBS, 3 x 3 minutes.
9. Stain with diaminobenzidin (DAB) solution 10 minutes at room temperature.
10. Wash with running tap water, 3 minutes.
11. Counterstain with Mayer's hematoxylin, 2 minutes.
12. Wash with running tap water, 5 minutes.
13. Dehydrate with increasing solution of ethanol; 50%, 70%, 96%, absolute, 3 minutes each.
14. Clear with xylol, 3 x 3 minutes.
15. Mount with mounting medium (e.g. malinol).

Indirect Immunoperoxidase Staining On Formalin-Fixed And Paraffin Embedded Tissues:
1. 4 micron thick sections should be used.
2. Dewax in Xylol, 3 x 3 minutes.
3. Rehydrate in decreasing grades of ethanol:absolute, 96%, 70%, 50%, 3 minutes each.
4. Block endogenous peroxidase activity with freshly made 0.3% H2O2 in methanol, 20 minutes.
5. Wash with PBS, 3 x 3 minutes.
Only if trypinsination is required
5a. Incubate sections with 0.1% Trypsin in 0.1% CaCl2 pH 7.6 for 10 minutes at room temperature.
5b. Wash with PBS, 3 x 3 minutes.
6. Cover the sections with 20% normal rabbit serum in PBS or normal human serum and incubate overnight in a humidity chamber at room temperature to reduce non specific background staining.
7. Decant 20% normal rabbit serum.
8. Incubate with monoclonal antibody (diluted in PBS), 1-2 hours at room temperature.
9. Wash with PBS, 3 x 3 minutes.
10. Incubate with peroxidase labeled second antibody, 30-60 minutes at room temperature.
11. Wash with PBS, 3 x 3 minutes.
12. Stain with diaminobensidin (DAB) solution, 10 minutes at room temperature. A stock solution of 0.5% DAB in 0.5 DAB in 0.5M Tris/HCl (pH7.4) can be made and stored frozen in the dark. Before use a quantity needed for staining can be thawed and diluted 10x with water. The diluted DAB solution should be filtrated. Just before use H2O2 must be added to a final concentration of 0.01%.
13. Wash with running tap water, 3 minutes.
14. Counterstain with Mayer's hematoxylin, 2 minutes.
15. Wash with running tap water, 2 minutes.
16. Dehydrate with increasing solutions of ethanol:50%, 70%, 96%, absolute, 3 minutes each.
17. Clear with xylol, 3 x 3 minutes.
18. Mount with mounting medium (e.g. malinol).
General Readings
  1. H.J. Beerstecher et al, 1988, J. of Histochemistry and Cytochemistry: vol. 36, 1153-1160.
Storage Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.
Avoid repeated freezing and thawing.
Shelf life: one year from despatch.
Format
Buffer System:
1% BSA and 20 mM Sodium Azide
State:
Liquid supernatant
Supernatant
Specificity
Specificity:
Reacts with EGF. In immunohistochemistry
positive staining is observed in salivary glands.
Species:
Mouse. Does not work in Rat.

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