Navigation Links
Use of ZipTip Pipette Tips for Microscale Affinity Mass Spectrometry

Overview

Affinity chromatography forms the basis of the separation and purification of a number of biological molecules. For example, antibodies may be purified based on their affinity for protein A, or a protein may be purified on the basis of its affinity for a monospecific antibody raised against it. Typically, these purifications take place in a chromatographic column with a relatively large volume. This note discusses the use of the C18 ZipTip pipette tips for conducting affinity-based separations on a microliter scale.

Key Features

Affinity Separation Protocol

Examples of ZipTip Pipette Tip Affinity Separations

Experimental Conditions

Accelerating Voltage: 25 kV Grid Voltage: 93%

Mode: Linear Guide Wire Voltage: 0.1 to 0.3%

Matrices: Sinapinic acid (SA) at 10 mg/mL in 50% MeCN/0.2% TFA, α-cyano-4-hydroxycinnamic acid at 10 mg/mL in 50% MeCN/0.2% TFA.

Affinity Separation Protocol

Wash ZipTip pipette tip (MeCN, 50% MeCN, 0.1% TFA)

Incubate ZipTip pipette tip with Affinity Agent, e.g. Ab

Wash with incubation buffer

Incubate ZipTip pipette tip with blocking agent, e.g. BSA

Wash with incubation buffer

Incubate with sample, e.g., antigen Wash with incubation buffer, then water

Elute sample with acid, e.g., 1% TFA Elute with 50% MeCN

We followed the affinity separation protocol above. After conditioning the ZipTip pipette tip (Figure 1), a goat anti-human transferrin was incubated with it by passing 20 μL of anti-serum up and down the tip ten times. After 3 washes with 10 μL aliquots of phosphate buffered saline at pH 7.4 (PBS), 20 μL of a 20 mg/mL solution of bovine serum al
'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Brinkmann/Eppendorf SOP Standard Operating Procedure for Pipettes
2. Highly Active and Removable AffinityTagged Lambda Phosphatase
3. Double-Affinity Purification of GST-His Tagged Proteins Using Vivapure 8-to-96 Well Cobalt-Chelate Kit
4. Affinity Depletion Cartridges for Removal of Human Serum Albumin and Immunoglobulins from Human Serum
5. Identification of Low-Level Oxidation Products in Calmodulin Using NanoSpray Ion Trap Mass Spectrometry
6. Determination of N-nitrosamines in Baby Bottle Rubber Teats by Liquid Chromatography-Atmospheric Pressure Chemical Ionization Mass Spectrometry
7. Small-Molecule Quantitation Using Automated Single-Shot Averaging MALDI-TOF Mass Spectrometry
8. Achieving Reliable Protein Quantitation Results Using ICAT Reagents, MALDITOF Mass Spectrometry, and Advanced Data Interpretation Tool
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Use ZipTip Pipette Tips for Microscale Affinity Mass Spectrometry

(Date:12/16/2009)... company,s revenues increase 22% for fiscal 2009; on ... , TORONTO, Dec. 16 /PRNewswire-FirstCall/ - Microbix Biosystems ... and biological technologies, today reported financial results for ... year ended September 30, 2009, Microbix reported revenues ... same period in 2008. For fourth-quarter 2009, the ...
(Date:12/16/2009)... Eppendorf has initiated an,import ban ... systems for,infringement of European patent EP 1 ... and/or the quantification of a target compound". ... of the respective products into any country ... to confiscate any,violating products. , ...
(Date:12/15/2009)... Institute (JBEI), one of three Department of Energy (DOE) ... service to protect its valuable intellectual property. , ... ... December 15, 2009 -- Surety, LLC , the ... today announced that the Joint BioEnergy Institute (JBEI), one ...
(Date:12/15/2009)... Medical, Inc. (OTC Bulletin Board : AEMD) announced ... Comper have joined the Science Advisory ... Schorey and Dr. Comper are leading researchers in ... of Aethlon Medical, was recently formed to leverage ... being advanced as an adjunct therapy for Hepatitis-C ...
Breaking Biology Technology:Microbix' Pipeline Set To Reach Milestones In 2010 2Microbix' Pipeline Set To Reach Milestones In 2010 3Microbix' Pipeline Set To Reach Milestones In 2010 4Microbix' Pipeline Set To Reach Milestones In 2010 5Microbix' Pipeline Set To Reach Milestones In 2010 6Microbix' Pipeline Set To Reach Milestones In 2010 7Microbix' Pipeline Set To Reach Milestones In 2010 8Eppendorf Successfully Initiates Import ban Against Nanosphere, Inc. for the Countries of the European Union 2Joint BioEnergy Institute Selects Surety's AbsoluteProof to Protect the Integrity and Legally Defend the Authenticity of its Scientific Intellectual Property 2Joint BioEnergy Institute Selects Surety's AbsoluteProof to Protect the Integrity and Legally Defend the Authenticity of its Scientific Intellectual Property 3Joint BioEnergy Institute Selects Surety's AbsoluteProof to Protect the Integrity and Legally Defend the Authenticity of its Scientific Intellectual Property 4Joint BioEnergy Institute Selects Surety's AbsoluteProof to Protect the Integrity and Legally Defend the Authenticity of its Scientific Intellectual Property 5Aethlon Medical Appoints Jeff Schorey and Wayne Comper to Exosome Sciences Advisory Board 2Aethlon Medical Appoints Jeff Schorey and Wayne Comper to Exosome Sciences Advisory Board 3Aethlon Medical Appoints Jeff Schorey and Wayne Comper to Exosome Sciences Advisory Board 4Aethlon Medical Appoints Jeff Schorey and Wayne Comper to Exosome Sciences Advisory Board 5
... ImQuest Life Sciences today presented important new results ... inhibitor of HIV-1 at the 16th CROI meeting ... the advanced development of ImQuest,s lead HIV pyrimidinedione ... of HIV, exhibiting a dual mechanism of action ...
... regulatory filings for S0812 and additional promising bitter ... Inc. (Nasdaq: SNMX ), a leading ... and develop novel flavor ingredients for the food, ... it has initiated development activities for S0812 and ...
... In a paper,published today, scientists at deCODE genetics ... continents present a novel insight into the connection,between ... in some,10,000 Icelanders, the deCODE team discovered several ... genome (SNPs) that increase levels of,eosinophils in the ...
Other Biology Technology:ImQuest Presents IND-Enabling Safety Pharmacology and Pharmacokinetic Data for the Therapeutic HIV Compound IQP-0410 at Conference on Retroviruses and Opportunistic Infections 2SENOMYX ANNOUNCES INITIATION OF DEVELOPMENT ACTIVITIES FOR NEW BITTER BLOCKERS 2SENOMYX ANNOUNCES INITIATION OF DEVELOPMENT ACTIVITIES FOR NEW BITTER BLOCKERS 3SENOMYX ANNOUNCES INITIATION OF DEVELOPMENT ACTIVITIES FOR NEW BITTER BLOCKERS 4Multinational deCODE Study Identifies New Link Between Inflammation and Heart Attack 2Multinational deCODE Study Identifies New Link Between Inflammation and Heart Attack 3Multinational deCODE Study Identifies New Link Between Inflammation and Heart Attack 4
(Date:12/16/2009)... molecules form an intricate network of interactions. Today,s ... individual molecular reactions outside the cells. Since molecular ... the laboratory, scientists suspect that the kinetics of ... external probes. We expected the cellular reaction speed ... Braun. "However, our novel optical approach showed that ...
(Date:12/16/2009)... Hill, CA Tom Bowman, an expert in communicating ... Global Change, has developed a series of graphics that ... Climate Change (IPCC) report for public audiences. These new ... they need to make informed decisions about climate change ... as a means to clearly and accurately communicate findings ...
(Date:12/16/2009)... -- DigitalPersona, Inc., a leader in fingerprint ... agreement with IBM to deploy a new ... on IBM SurePOS 500 retail systems. The ... to more easily and quickly identify users ... to the individuals who perform them. ...
Breaking Biology News(10 mins):Looking for the heartbeat of cellular networks 2Bowman creates graphic translation of climate change data 2DigitalPersona Fingerprint Sensor Technology Ships on IBM SurePOS 500 Retail Systems 2Senators Gregg and Shaheen Vote to Protect Americas Kids From Tobacco 48699 1Senators Gregg and Shaheen Vote to Protect Americas Kids From Tobacco 48699 2Senator Bill Nelson Votes to Protect Americas Kids from Tobacco 48696 1Senator Bill Nelson Votes to Protect Americas Kids from Tobacco 48696 2Senators Specter and Casey Vote to Protect Americas Kids from Tobacco 48693 1Senators Specter and Casey Vote to Protect Americas Kids from Tobacco 48693 2
... chicken when she crossed the road is less important that what ... , Organic arsenic is fed to poultry to prevent bacterial infections ... up by the tissue and the majority of it is excreted ... urine from under poultry houses -- is rich in nitrogen and ...
... prediction of patient outcomes, lead to tailored treatmentsWhen it ... can be deadly. In some severely injured patients, this ... response, including bloodstream infection (sepsis) and multiple organ failure. ... is still mysterious, so doctors can't accurately predict how ...
... Russia, not the least of which is a very ... 25- to 54-year-old Russian men living in an industrial ... "surrogate" alcohols, otherwise known as products containing alcohol that ... now analyzed the contents of these surrogate alcohol products, ...
Other Biology News:New class of antibiotics effective against drug-resistant bacteria discovered in fungi 2Study Unlocks Insects?Secret for Love in the Dark 2Study Unlocks Insects?Secret for Love in the Dark 3Russian 'surrogate' alcohols are a killer 2Russian 'surrogate' alcohols are a killer 3Russian 'surrogate' alcohols are a killer 4
Griess Reagent component for determination of nitrite in aqueous solutions....
Human HVEM/TNFRSF14 Allophycocyanin MAb (Clone 94801)...
LAL Chromogenic Endpoint assay...
RayBio Human Atherosclerosis Antibody Array 1.1 (4) with Accessory, detects 40 inflammatory factors (for lysate) Class: Antibody Array Products Product Group: Antibody Array...
Biology Products: