Hi Gabriela,

Try GE Amersham!

Gelfiltration:

http://www.gelifesciences.com/webapp/wcs/stores/servlet/catalog/de/GELifeSciences/products/AlternativeProductStructure_17393/

Ion Exchange:

http://www.gelifesciences.com/webapp/wcs/stores/servlet/catalog/de/GELifeSciences/products/AlternativeProductStructure_17442/

Cheers

Ignazio

Hi Gabriela,

Try GE Amersham!

Gelfiltration:

http://www.gelifesciences.com/webapp/wcs/stores/servlet/catalog/de/GELifeSciences/products/AlternativeProductStructure_17393/

Ion Exchange:

http://www.gelifesciences.com/webapp/wcs/stores/servlet/catalog/de/GELifeSciences/products/AlternativeProductStructure_17442/

Cheers

Ignazio

Thanks Ignazio!

Good suggestions!

About ION EXCHANGE, I warmly suggest you to try also Merck resins "Fractogel":

http://www.merckmillipore.it/pharmaceutical-ingredients/fractogel-/c_XRmb.s1OY_sAAAEoDPY71veP?PortalCatalogID=merck4pharma&CountryName=Italy

I did comparative studies with different types of resins (of course GE), but the Fractogel resulted the best in absolute!

Another choice (very good, almost Pressure and flow unaffected) are the products from BiaSeparations:

http://www.biaseparations.com/pr/1129/cim-so3-1-tube-monolithic-column

About Gel Filtration, I agree with Ignazio: GE the best.

Good luck,

Barbara

We use GE columns on both BioLogic and Akta systems, it works very well !

Check GE Healthcare, although you may need to buy an adaptor also

Our lab has been using GE columns for gel filteration. We have superdex 200 16/60 which have been working really well but small column superdex 200 10/300 gave lots of problem especially with when buffer had 5% glycerol. It gets choked easily. So, it depends whether you want to see just the profile of the protein or you want to get the purified protein, you may choose the column size on the basis of your requirement. I have no idea about ion exchange .

For size exclusion chromatography (=gel filtration) we use Toyopearl HW-55S resin with full satisfaction (more see http://www.separations.us.tosohbioscience.com/Products/ProcessMedia/ByMode/SEC/ToyopearlHW55.htm). For any type of ion exchange chromatography (strong and/or weak, anion and/or cation exchange ...) we use BioRad resins with full satisfaction as well. However, you are right to try different resins when you plan to establish a purification protocol for the separation of your particular protein.

we also have had good experience with GE, some help in selecting the right IEX for you http://wolfson.huji.ac.il/purification/PDF/IonExchange/GE_IEXcolumns.pdf and SEC http://kirschner.med.harvard.edu/files/protocols/GE_gelfiltration.pdf

we also work since many years with the GE columns. But the choice depend on the protein(s) you want to purify.

Regards

For conventional anionic chromatography we used successfully DEAE-Toyo-Pearl (from Toyo-Soda). This one has high capacity, fast, doesn't swell and resistant to microflora. Filtration gels from Toyo have broad spectrum of mol.mass of substances for separation and are really good.

For anion exchange column (weak or high) I buy the supports from Biorad and I do pack my own columns for preparative purifications, for GF I prefere the supports or columns from Amersham (GE Healthcare) they have good choise for different separation ranges.

With respect to hardware, the DuoFlow uses 1/4"-28 fittings, while the old-style GE (Pharmacia) columns are M6 and the Tricorn series are 1/16" fingertight. See

www.bio-rad.com/webroot/web/pdf/lsr/literature/Bulletin_5326A.pdf‎

for a conversion chart and part numbers.

As for media (or prepacked columns), I have routinely used GE columns and media with a DuoFlow. Exactly what media you want will depend on your application (e.g., you probably don't want to put lysate over a Source15), and you might need more than one of each type (anion/cation/SE) - I have Q and S Source15 and Fast-Flow, a half-dozen or so size exclusion columns, and an assorted collection of affinity resins. Some are in Bio-Rad econo-columns with flow adapters, most are in GE XK or Tricorn. All are driven by a DuoFlow.

The GoPure columns with POROS media are great for ion exchange, they are 5-10-fold faster than similar medias from GE (S HP and Q HP) because of the very low back pressure, durable with a high binding capacity, and very reasonably priced for a pre-packed "non-disposable" column.

https://www.lifetechnologies.com/search/global/searchAction.action?query=Gopure&resultPage=1&resultsPerPage=15

Go ahead with GE product. Just make sure you know your protein well (e,g pI and MW) before you place your order. The types of ion-exchange i.e Mono Q/S, Resource Q/S, HP series etc is depend on what you intended to do. If ion exchange is for capturing or fractionation, use HP series. Resource/Mono/Mini Q-S is powerful for polishing (better resolution).

Hi Gabriela,

I have used GE, BioRad and Toyo columns with Biologic DuoFlows and AKTA FPLCs. These manufacturers will have columns that suit your needs and you can buy adaptors to fit any column to any FPLC.

Personally I have been trying to move away from GE as they are expensive and not very willing to bargain on prices. Their columns are good though! The Toyo columns are often cheaper and still give good separation albeit slightly different from my experience... I think due to chemistry... but if you are starting the purification from scratch then this wont matter too much.

If you are going outside those three... the biggest thing you need to watch out for is the resin type as some resins (silica based?) can not handle pH 7.5 and above as they start to degrade. Just ask the sales rep and they should be able to tell you.

Russell

Thanks everyone for your help.

1 GE Healthcare

IEX:

Q Sepharose™ FF

Q Sepharose™ HP

Q Sepharose™ BB

Q Sepharose™ XL

DEAE Sepharose™ FF

ANX Sepharose™ 4FF low sub

ANX Sepharose™ 4FF high sub

SOURCE™ 15Q

SOURCE™ 30Q

Capto™ Q

Capto™ DEAE

Capto™ Q ImpRes

SP Sepharose™ FF

SP Sepharose™ HP

SP Sepharose™ BB

SP Sepharose™ XL

CM Sepharose™ FF

CM Sepharose™ HP

SOURCE™ 15S

SOURCE™ 30S

Capto™ S

MacroCap SP

Capto™ SP ImpRes

GFC:

Sephadex™ G-10

Sephadex™ G-15

Sephadex™ G-25 Coarse

Sephadex™ G-25 medium

Sephadex™ G-25 Fine

Sephadex™ G-25 Superfine

Sephadex™ G-50 Coarse

Sephadex™ G-50 medium

Sephadex™ G-50 Fine

Sephadex™ G-50 Superfine

Sephadex™ G-75

Sephacryl S-100 HR

Sephacryl S-200 HR

Sephacryl S-300 HR

Sephacryl S-400 HR

Sephacryl S-500 HR

Superdex™ 200 prep grade

2 Merck Millipore

IEX:

Fractogel® EMD DMAE (M)

Fractogel® EMD DEAE (M)

Fractogel® EMD DEAE (S)

Fractogel® EMD TMAE (M)

Fractogel® EMD TMAE (S)

Fractogel® EMD TMAE Hicap (M)

Fractogel® EMD TMAE Medcap (M)

Eshmuno™ Q

Fractogel® EMD COO- (M)

Fractogel® EMD COO- (S)

Fractogel® EMD SO3- (M)

Fractogel® EMD SO3- (S)

Fractogel® EMD SE Hicap (M)

Eshmuno™ S

Eshmuno® CPX

3 Pall Corporation

IEX:

Q HyperCel™

DEAE Ceramic HyperD® F

Q Ceramic HyperD® 20

Q Ceramic HyperD® F

HyperCel™ STAR AX

S HyperCel™

CM Ceramic HyperD® F

S Ceramic HyperD® 20

S Ceramic HyperD® F

4 Bio-Rad Laboratories

IEX :

Macro-Prep® DEAE

Macro-Prep® High Q

Macro-Prep® 25 Q

UNOsphere™ Q

Nuvia™ Q

Nuvia™ cPrime™

Macro-Prep® High S

Macro-Prep® 25 S

Macro-Prep® CM

UNOsphere™ S

UNOsphere™ Rapid S

Nuvia™ S

5 Life Technologies

IEX:

POROS® 50 HQ

POROS® 50 PI

POROS® 50 D

POROS® 20 HQ

POROS® 50 HS

POROS® 20 HS

POROS® XS

6 Avantor Performance Materials

IEX : Multimode

BAKERBOND™ POLYQUAT

BAKERBOND™ POLYPEI

BAKERBOND™ POLYCSX

BAKERBOND™ POLYABx

7 YMC

IEX:

YMC - BioPro Q30

YMC - BioPro Q75

YMC - BioPro S30

YMC - BioPro S75

8 Tosoh Bioscience

IEX:

Toyopearl® DEAE-650C

Toyopearl® DEAE-650M

Toyopearl® DEAE-650S

Toyopearl® SuperQ-650C

Toyopearl® SuperQ-650M

Toyopearl® SuperQ-650S

Toyopearl® QAE-550C

Toyopearl® GigaCap® Q-650M

Toyopearl® Q-600C AR

TSKgel® SuperQ-5PW (20 µm)

TSKgel® SuperQ-5PW (30 µm)

Toyopearl® CM-650C

Toyopearl® CM-650M

Toyopearl® CM-650S

Toyopearl® SP-650C

Toyopearl® SP-650M

Toyopearl® SP-650S

Toyopearl® SP-550C

Toyopearl® MegaCap® II SP-550 EC

Toyopearl® GigaCap® S-650M

Toyopearl® GigaCap® CM-650M

GFC:

Toyopearl® HW 40S

Toyopearl® HW 40F

Toyopearl® HW 40C

Toyopearl® HW 50S

Toyopearl® HW 50F

Toyopearl® HW 55S

Toyopearl® HW 55F

Toyopearl® HW 65S

Toyopearl® HW 65F

Toyopearl® HW 65C

Toyopearl® HW 75F

9 Chisso Corporation

IEX:

Cellufine® MAX Q-r

Cellufine® MAX Q-h

Cellufine® Q-500 (m)

Cellufine® MAX S-r

Cellufine® MAX S-h

Cellufine® C-500 (m)

10 Resindion

IEX:

ReliSorb™ QA400/SS

ReliSorb™ DA400/SS

ReliSorb™ SP400/SS

ReliSorb™ CM400/SS

11 Agilent Technologies

PL-SAX 1000Å 30µm

PL-SAX 1000Å 10µm

PL-SCX 1000Å 30µm

PL-SCX 1000Å 10µm

12 ProMetic Life Sciences

DEAE PuraBead HF

Best Luck.

Write me to [email protected] if you need more information.

This is an old thread, but anyway.... We frequently do routine ion-exchange bind-release cleanups in eppendorves in batch mode, without an FPLC machine (like one would do an IP). GE sepharose resins work quite well for this, and one can get 50 ml of beads for 1/3 of a price of a prepacked 10ml column (or even cheaper) Batch mode works very well for smal volumes (e.g. 200 ul), and one can do several samples in parallel. Recommended for well-chracterized proteins whose elution profiles are known.