说明书：

MF081- M5 GFP-Tag(7G9)mAb.pdf

【STORAGE】

Store at -20°C. Stable for one year from the date of shipment.

【BACKGROUND】

The green fluorescent protein (GFP) was originally identified as a protein involved in the bioluminescence of the jellyfish Aequorea victoria. GFP cDNA produces a fluorescent product when expressed in prokaryotic cells, without the need for exogenous substrates or cofactors, making GFP a useful tool for monitoring gene expression and protein localization in vivo. Several GFP mutants have been developed, including EGFP, which fluoresce more intensely than the wildtype GFP and have shifted excitation maxima, making them useful for FACS and fluorescence microscopy as well as double-labeling applications. GFP is widely used in expression vectors as a fusion protein tag, allowing expression and monitoring of heterologous proteins fused to GFP.

【REFERENCES】

1.Prasher, D.C., et al. 1992. Primary structure of the Aequorea victoria green fluorescent protein. Gene 111: 229-233.

2.Chalfie, M., et al. 1994. Green fluorescent protein as a marker for gene expression. Science 263: 802-805.

【SOURCE】

This monoclonal antibody is produced by immunizing animals with full length recombinant GFP.

【SPECIFICITY】

GFP-Tag (7G9) Mouse mAb detects GFP and GFP fusion proteins.

【REACTIVITY】

All

【ISOTYPE】

Mouse IgG1

IMPORTANT

Use an anti-MOUSE secondary antibody to detect the 7G9 antibody.

RECOMMENDED ANTIBODY DILUTIONS

Western blotting            1:5000

Immunofluorescence        1:500

Immunoprecipitation         1:50

ELISA                     1:2000

For Western blots, incubate membrane with diluted antibody in 5﹪w/v nonfat dry milk,1X tbs,0.05﹪Tween-20 at 4˚C with gentle shaking,overnight.

APPLICATION DATA

Western blot analysis of over-expressed GFP-tagged protein in 293T cell lysate,using M5 GFP-tag (7G9) Mouse mAb.The antibody dilutions are 1:5000(lane1)and1:10000(lane2).Each lane was loaded with 10μg of cell lysate.

IP of extracts from 293T cells transfected (lane1,2)with a GFP-tagged protein and a mock transfection (lane3,using the same protein without the GFP tag),using M5 GFP-Tag(7G9) mouse mAb and probed on Western biot using the same antibody. Dilution: 1:50(1),1:100(2),1:50(3).

IF analysis of 293T cells transfected with a GFP-tagged protein, using M5 GFP-Tag (7G9) Mouse mAb at a 1:500 dilution.