YOSHIDA BIOLOGICAL LABOLATRY・About the RFHR metod of 2D PAGE

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Protein analysis services by the RFHR 2D PAGE

Protein analysis services by the RFHR 2D PAGE

No isoelectric point method is applied to the RFHR 2D PAGE, and instead, definite pH electrophoresis are carried out at the first dimension (pH 8.6 or 9.8) and the second dimension (pH 3.4).Prior to the first dimension electrophoresis, "the 0th dimension electrophoresis " is introduced for high concentration of samples.In the RFHR 2D PAGE, all proteins are separated by the difference of the migration rate but not the isoelectric point even at the first dimension. Therefore this method has a high separation ability for basic proteins.

RFHR 2D PAGE has a high identification rate for protein genes from protein spots on the 2D gel, as protein capacity of the gels is very high (5mg/gel). The RFHR 2D PAGE can avoid artificial splitting of protein spots caused by isoelectric point method, and can determine the number of each protein in a cell by densitometry of the protein spot on the 2D gel. The RFHR 2D PAGE is suited for studies on post-translational modification and analyses of fine proteins, as it has no artifact to split protein spots. The RFHR 2D PAGE is suited for functional analysis of proteins after electrophoresis, as this method uses no SDS.

[Topics of the RFHR]

Escherichia coli ribosomal proteins (1986)

E. coli ribosome is a big particle which consists of 3 species of RNAs and 54 species of proteins.Iso-electric point electrophoresis (IPG method) cannot separate ribosomal priteins well, because about 90% of ribosomal proteins are basic.On the contrary the RFHR electrophoresis can separate enough and quantitatively.In 1986, ribosomal protein L35(A) and L36(B) were detected and identified by the RFHR method, and at last definition of E. coli ribosomal proteins was completed.

Weekly acidic proteins involved in E. coli soluble fraction

The RFHR method has a highly quantitative separation ability to most proteins without discrimination of iso-electric point. Molecular number of each protein in a cell can be determined by densitometry of protein spots on the RFHR 2D gels.

YOSHIDA BIOLOGICAL LABORATORY INC.
analyzes your protein samples by the RFHR 2D PAGE.

The contents of analysis：
・Offer your desalted and lyophilized protein samples to us.
・However, we can prepare your protein samples including desalting and lyophilization prior to RFHR 2D PAGE, if you hope it.
・We carry out a pretest whether your sample is suit for RFHR 2D PAGE or not, and if it is not suit, we require the re-preparation.
If the re-preparation fails, we stop the analysis. In this case, the fee for pretestis necessary to you.
・We analyze your protein samples by the RFHR 2D PAGE, and send the results of the spots pattern on the 2D gel.
・We can accept your orders for identification of the proteins as well.