High Resolution Accurate Mass Spectrometer System (HRAMS) For Proteomics and Metabolomics

Required Samples :-

Guidelines for Sample Preparation

For Proteomics

• Prevent or minimize contamination of your sample. Contaminants can be keratins, plasticizers, polymers, detergents, and non-volatile buffers.
• Fungal contamination in protein gels is very common, avoid such types of gels for processing.

Protein storage recommendations – Proteins in coomassie-stained gels are relatively stable at 4? or -20?. Silver-stained protein bands must be analysed as soon as possible after it is isolated and purified or should be kept frozen (i.e. -20? or -80?).

Protein solution form - must be stored and transported at -20? or -80?.

For In-gel analysis- Coomassie stain are preferred and will generally result in the best peptide recoveries and protein ID results.

• Cut the band out as close as possible; be very selective and cut out only the centre of the band; don’t include any excess gel material.
• Purity of the protein sample is critical for a successful analysis of the intact molecular weight and identification of a protein.

For In-solution analysis-

• For In solution analysis the amount of protein must be 100- 150 μg for each sample.
• It is critical that the sample does not contain any detergents, Surfactants, or polymers (even at trace levels).
• Avoid using Triton, Tween, SDS, and PEG, similar to that during any part of the protein preparation.
• Use HPLC/FPLC to purify and collect samples whenever possible.
• For more than 10 samples new requisition form needs to be submitted.

General Instruction:

• Sample will not be analysed till the payment confirmation.
• Before sample preparation, it is advisable to contact the facility and discuss the protocol.

Disclaimer: The experimental data provided is only for research & development purposes. These cannot be used as a certificate in legal disputes.

For Metabolomics

• The metabolites should be extracted by the user itself in the proper solvent of good quality (LC-MS or HPLC grade).
• The sample should be pure and dried and submitted in a properly labeled vial
• Mention the suitable solvent (preferably protonated solvent like MeOH, ACN, or Water.)

Note:

• The users are requested to contact the facility in case of queries regarding the price and analysis to be used. Email of the facility: sathiservices@bhu.ac.in.
• Sample list in Excel sheet must be provided to the facility with the weight of samples and the sample tubes must have similar codes for ease of handling (Lab name- day and month- sample number. Eg. JV-2611-01).
• All the samples and standards must be prepared by using MS grade solvents for a good quality result.
• Average processing time after sample submission is 3 weeks for metabolites and 4-8 weeks for proteomics. The sample processing time starts after the submitted samples.
• After analysis the raw data will be provided without any statistical processing.
• Facility will not be responsible for data processing/experimental results.
• Kindly acknowledge the facility in your publications.

**Please make available the analysis related publications to expedite the sample preparation related protocols.

Form Application :-

User Charges :-

In Case of Deep Protein Analysis :-

For Proteomics Quantitative analysis Rs. 1000/- per sample additional charges will be applicable.

In case of Single Metabolite Identification HRMS with database search and Total Metabolite Profiling HRMS with database searchMetabolite Identification and Profiling :-

• If the run time exceeds the limit the charges will be in multiple accordingly.
• These charges are for the both (positive + negative) modes.
• For single (positive + negative) mode small molecules identification the charges will be reduced accordingly.
• For Quantitative analysis of small molecule Rs. 1000/- per sample additional charges will be applicable.

Note – For External samples, GST will be applicable as per rule.

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