BM2311 Melanoma Associated Antigen 450 / 250kDa antibody

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Mouse anti Human Melanoma Associated Antigen NKI/M6


Product Description for Melanoma Associated Antigen

Mouse anti Human Melanoma Associated Antigen NKI/M6.
Properties: 450 / 250kDa
Presentation: Purified
Product is tested for Frozen Sections.

Properties for Melanoma Associated Antigen

Product Category Primary Antibodies
Quantity 1 ml
Presentation Purified
Reactivity Hu
Applications C
Clonality Monoclonal
Clone NKI/M6
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

Property 450 / 250kDa
Isotype control AM03095PU-N, SM10P (for use in human samples)
Application Suitable for use in staining of Melanoma Cells in frozen sections by regular immunoperoxidase and Immunofluorescence tests. A 1:5-1:10 dilution in phosphate buffered saline is suggested.
For further instruction, see A and I in ''Protocols''.
Concentration 0.1 mg/ml

Membrane Immunofluorescence:
1. Transfer 5x10e5 cell in 25 µl to a 10x75 mm test tube for staining.
2. Add 25-50 µl of the diluted monoclonal antibody, mix and incubate for 30 minutes on ice.
3. Wash twice with PBS containing 1% BSA and 0.02% Sodium Azide.
4. Add 50 µl of appropriate FITC/TRITC labeled anti mouse Ig antibody, mix and incubate for 30 minutes on ice.
5. Wash twice with PBS containing 1% BSA and 0.02% Sodium Azide.
6. Resuspend the cells in 100 µl washing buffer and analyze on a fluorescence activated cell sorter (FACS) or mount in 90% glycerol, 10% 1 M Tris/HC1, pH 8.0 and analyze by fluorescence microscopy.


Immunoperoxidase Test On Sections:
1. Frozen sections should have been fixed in acetone for 10 min.
2. Incubation in antisera 40-60 min.
3. Incubation in conjugate (e.g. peroxidase conjugated anti mouse IgG), 30 min.
4. Wash in PBS 2 X 5 min.
5. Incubation in AEC, 0.0.1% H2O2, 10 min.
Preparation substrate: 5 mg AEC (3-amino-9-ethylcarbozole) is solubilized in 0.5 ml DMF (dimethylformamide). A glass (or acetone resistant plastic) tube or pipet should be used!
add 9.5 ml. 0.05M NaAc buffer, pH 4.9.
add 5 µl 30% H2O2.

6. Wash in demi water, 2 x 5 min.
7. Slightly counterstain in hematoxylin e.g. 10 sec.
8. Wash in tap water until sections are blue.
9. Mount in aquamont and examin.
General Readings
  1. de Vries, J.E., et. al., 1986, Int. J. Cancer, 38, 465.
  2. Natali P.G., J. Nat'l. Cancer Inst. 67, 591.
  3. Buraggi, G.L., et al., 1985, Cancer Res., 45, 3378.
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.
Buffer System:
PBS with 0.1% BSA as stabilizer and 0.09% Sodium Azide as preservative.
Liquid purified Ig fraction.
Recognizes a high molecular weight proteoglycan with a molecular weight of > 450 kD (chondroitin sulfate) and 250 kD (core protein).
Reacts strongly with Melanoma Cells derived from cell lines and short term cultures and reacts preferentially with Melanoma Cells in frozen tissue sections. (1)
Reacts with most nevi and perineurium and shows weak reactivity with hair follicles.

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