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蛋白質翻譯后修飾的驗證
發布時間:2018-4-2 11:14:10 信息來源: 藍光生物 編輯者:藍光生物 瀏覽次數:1446

Issues in Validation of    Post-Translational Modifications.

Why are proteins, detected by    mass spectrometry, not validated by site-specific antibodies?

The modified motif could be detected by mass spectrometry    (MS) after affinity enrichment, then the modified protein should be    affinity-enriched for western-blotting validation, ELISA analysis with the    the sequence-specific, modified peptide antibodies.

Can the sequece-specific modifed    peptide antibody used for affinity-enrichment?

Yes!

Only if this modified peptide antibody is able to    immunoprecipitation (IP) the modified proteins.


   

No!

Not all the peptide antibodies are able to perform IP    studies, even if it could be utilized for Western blotting (WB).

By using the pan-specific modified amino acid antibodies to    affinity-enrich all the modified proteins, you may selectively detect the    target modified proteins by WB using modified sequence-specific antibodies.

For WB, you could only resolve maximum 100 ug/lane of crude    proteins in SDS-PAGE, your target modified proteins in 100 ug of crude    lysates may not be detectable and may also interfere with other abundant    proteins, but you could enrich  your target modified proteins only    from 1-10 mg of crude proteins, the enriched modified proteins could    readily be resolved in SDS-PAGE and become detectable level for WB.

Application Examples:

Three trimethylated sites of histone H3 enriched with 100    uL of the pan trimethyl lysine specific antibodies (ICP0610) Agarose (1 mg    of the antibodies) from 5 mg crude proteins, detected by WB with three    site-specific antibodies.

PTM Proteomic Analysis Sample Preparation Scheme

Follow our schematic for consistent and accurateresults.


Biotin Antibody Agarose -        ICP0615

Biotin is a vitamin, which binds strongly and        irreversibly to avidin. Biotin is commonly used for labelling reagents.        Anti-biotin antibody binds to biotin strongly but reversibly, the        binding could be competitively eluted with free biotin.

10mg - $695

Biotin Antibody, HRP -        ICP0612

This antibody has been used for direct competitive        ELISA for quantization of d-biotin and western-blot analysis of        biotinylated antibodies and proteins. The antibody reacts with free        biotin and biotinylated antibodies and proteins.

1mg - $325

Biotin Antibody, FITC -        ICP0613

This antibody was conjugated with FITC and has been        used for indirect fluorescent staining studies. It reacts with free        biotin and biotinylated proteins regardless of species.

1mg - $325

Biotinylated BSA - ICP0614

The biotinylated BSA is well recognized and binds        strongly to the affinity purified anti-biotin antibodies.

5mg - $275

Acetyl Lysine Antibody,        Agarose - ICP0388

Protein acetylation is a form of post-translational        modification known to regulate many diverse biological processes.        Affinity chromatography is one of the most efficient and rapid methods        to enrich and purify the acetylated species for further MS/MS        identification.

2mg - $550

5mg - $950

Methyl Lysine Antibody,        Agarose - ICP0588

Protein methylation is a form of post-translational        modification known to regulate many diverse biological processes.        Affinity chromatography is one of the most efficient methods to enrich        and purify the methylated species for further MS/MS identification.

5mg - $895

Trimethyl Lysine Antibody,        Agarose - ICP0606

Rapid isolation and purification of the trimethylated        species for proteomic profiling of the trimethylated proteins in        different stages of development, disease, and signal stimulation, and        identification of the protein trimethylation site.

5mg - $650

Phospho-Tyrosine Antibody,        Agarose - ICP9804

This tool is essential for proteomic profiling of the        phosphorylated proteins in different stages of development, disease,        and signal transduction, as well as identifying the precise site of        phosphorylation in a phospho-protein.

5mg - $650

Ochratoxin A Antibody,        Agarose - 1220

The Ochratoxin A antibody agarose is useful matrix for        affinity extraction and enrichment of OA from complex analytical        samples.

20mg - $550

Protein A Agarose - ICP1001

The immobilized protein A on the beaded agarose could        be utilized for the immunoprecipitation and purification of both        polyclonal and monoclonal antibodies from mammalian species. Protein A        is very heat and acid stable.

2mg - $150


 
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