PureProteome Protein A和Protein G磁性珠粒体数据表说明书

Data Sheet

Data Sheet

PureProteome

™

Protein A and

Protein G Magnetic Beads

Eliminate Variability. Maximize Recovery.

PureProteome Protein A and Protein G

magnetic beads are a powerful system to

isolate antibodies faster, easier, and with more

reproducibility then ever before. PureProteome

Protein A and Protein G magnetic beads purify

your sample with the highest binding capacity of

any magnetic beads available. You can achieve

reproducible results in both immunoprecipitation

and serum depletion assays.

Advantages

• High capacity: More than 6x the

binding capacity of competitive

magnetic beads.

– Bind 1.5-3.5 µg of rabbit IgG

per µL of suspension

• Consistent results: Total removal

of buffers with no sample loss

• Fast processing time: Bead

immobilization occurs in seconds

• Ideal for immunoprecipitation and

serum depletion assays

• Economical: Up to half the price of

competitive magnetic beads

Superior magnetic bead performance

from the experts in porous media

HigH BinDing CApACity

witH RApiD SAMplE pRoCESSing

Trust Millipore to develop a superior magnetic bead system

optimized to advance your research. Millipore’s process for

paramagnetizing porous silica particles enables us to provide

magnetic beads with the highest binding capacity when

compared to other magnetic purification systems.

With a binding capacity 6x greater then competitive beads,

PureProteome Protein A and Protein G magnetic beads

provide the greatest percentage of immunoglobulin (IgG)

depletion from serum samples with low non-specific binding.

get up to 8X greater depletion in half the time.

% lgG Depletion from 50 µL of Rabbit Serum Depletion Profile

Rabbit Serum (50 µL) diluted with PBS was incubated with

Protein A or Protein G magnetic beads per manufacturer’s

instructions. An ELISA assay was used to determine the

percentage of IgG depleted and the results were normalized by

the volume of suspension used per reaction.

Rabbit Serum (50 µL) diluted with PBS was incubated with

Protein G magnetic beads per manufacturer’s instructions. The

depleted rabbit serum samples were separated by SDS-PAGE

and the gel stained with Coomassie Blue using Millipore and

competitor magnetic beads. Lane 1: input material, lanes 2, 4, 6,

8: depleted samples, lanes 3, 5, 7, 9: eluted samples.

Use of the Indirect Immunoprecipitation Protocol

to Isolate p53 Protein from A431 Cell Lysates

A whole-cell lysate (300 mg of protein in 500 µL) prepared

from A431 cells was incubated in the presence of 5 µg of a

mouse monoclonal antibody specific for the tumor suppressor

protein/transcription factor p53 for 2 hours at 4 °C with

gentle mixing. The antigen-antibody complexes were then

incubated for 10 minutes at room temperature with 50 µL of

pre-washed PureProteome Protein A magnetic beads followed

by washes with PBS to remove any unbound protein. The

immunoprecipitated p53 protein was eluted from the beads

by incubation in the presence of gel loading buffer for 10

minutes at 70 °C. Ten microliter samples of the cell lysate

starting material (S), cell lysate that had been incubated in the

presence of the PureProteome magnetic beads (FT) and the

p53-containing eluted fraction (E) were subjected to SDS-

PAGE and western blot analysis using Immobilon-P blotting

membrane. The immunoreactive p53 protein was visualized

using the SNAP i.d.

™

Protein Detection System (antibodies:

rabbit anti-p53, HRP-conjugated goat anti-rabbit) and Immobilon

HRP Western Substrate. The results shown demonstrate

quantitative precipitation of the p53 protein from the samples.

2

CoMplEtE RECoVERy witH

REpRoDuCiBlE RESultS

Traditional PurificationMagnetic Bead Purification

Millipore’s new PureProteome magnetic beads

ensure the rapid and reproducible isolation

of proteins. Unlike conventional methods

that require centrifugation to pellet followed

by careful aspiration to avoid sample loss,

PureProteome magnetic beads are isolated

using a magnetic rack. This allows for the total

removal of buffers for complete recovery of

beads with no sample dilution.

Eliminate variability

while maximizing recovery.

Centrification toCareful removal of A magnetic field captures beads

pellet samplesupernatant is required against the tube wall allowing

to avoid sample losssupernatant to be removed easily

with no sample loss.

FASt AnD EASy

While traditional methods require minutes of harsh centrifuga-

Immunoprecipitation reactions that may require up to 18 hours

tion to isolate your sample, the PureProteome magnetic bead

of incubation with beads using conventional methods can now

system isolates proteins in seconds using a magnetic rack right

be accomplished in as little as 10 minutes.

on your bench. A magnetic field gently immobilizes the highly-

visible beads on the side of the tube. This allows quick and easy

Dramatically reduce your sample preparation

time with pureproteome magnetic beads.

aspiration and eliminates the need for a centrifugation step.

With the increased reaction kinetics of the Pure Proteome

magnetic beads, incubations can be performed in minutes.

RApiD DownStREAM pRoCESSing

To further speed up your immunodetection protocol,

detect your immunoprecipitated proteins with the

immunoprecipitation

SNAP i.d. Protein Detection System. Unlike conventional

western blotting, where diffusion is the primary means

protein Concentration

of reagent transport, this system applies a vacuum to

actively drive reagents through the membrane.

Electrophoresis

use pureproteome magnetic beads with

the SnAp i.d. System to isolate and detect

proteins in record time.

Membrane transfer

Blocking

Antibody Addition

Detection

3

oRDERing inFoRMAtion

DescriptionQty/pkCatalogue no.

DescriptionQty/pkCatalogue no.

PureProteome Magnetic Beads

PureProteome Protein A

Magnetic Beads

PureProteome Protein G

Magnetic Beads

Pure Proteome Nickel

Magnetic Beads

Magna GrIP

™

Rack

DescriptionnMwl

*

Qty/pk

Western Blotting

10 mL

2 x 1 mL

10 mL

2 x 1 mL

10 mL

2 x 1 mL

LSKMAGA10

LSKMAGA02

LSKMAGG10

LSKMAGG02

LSKMAGH10

LSKMAGH02

20-400

Catalogue no.

SNAP i.d.

™

Protein Detection System

Includes hardware base, blot holder

sample pack, blot roller, vacuum tubing

user guide.

DescriptionSize

1 kitWBAVDBASE

Qty/pkCatalogue no.

Immobilon

®

-P Transfer membrane (0.45 µm)

Cut Sheet8 x 10 cm

10 x 10 cm

20 x 20 cm

Roll

Cut Sheet

26.5 x 375 cm

8 x 10 cm

10 x 10 cm

20 x 20 cm

10

10

10

1

10

10

10

1

10

1

20

20

Qty/pk

IPVH08100

IPVH10100

IPVH20200

IPVH00010

ISEQ08100

ISEQ10100

ISEQ20200

ISEQ00010

IPFL10100

IPFL00010

IPSN07852

IPSN08132

Catalogue no.

Protein Concentration (< 4 mL samples)

Amicon

®

Ultra-4

Centrifugal Filter Unit with

Ultracel

®

-3 membrane

Amicon Ultra-4 Centrifugal

Filter Unit with Ultracel-10

membrane

Amicon Ultra-4 Centrifugal

Filter Unit with Ultracel-30

membrane

Amicon Ultra-4 Centrifugal

Filter Unit with Ultracel-50

membrane

Amicon Ultra-4 Centrifugal

Filter Unit with Ultracel-100

membrane

324UFC800324

Immobilon-PSQ Transfer membrane (0.2 µm)

1024UFC801024

3024UFC803024

Roll

Cut Sheet

Roll

26.5 x 375 cm

10 x 10 cm

26.5 x 3.75 cm

7 x 8.4 cm

8.5 x 13 cm

Immobilon-FL Transfer membrane (0.45 µm)

5024UFC805024

10024UFC810024

Blotting Sandwiches

Immobilon-P

Blotting Sandwich

* Nominal Molecular Weight Limit or membrane cut-off in kDa

Description

Western Blot Detection Substrates

Immobilon Western

Chemiluminescent HRP Substrate

Immobilon Western

Chemiluminescent AP Substrate

Spray & Glow

™

ECL Western Blotting

Detection System

Visit for additional pack sizes

100 mL

100 mL

100 mL

WBKLS0100

WBKDS0100

17-373

to plACE An oRDER oR RECEiVE

tECHniCAl ASSiStAnCE

Visit: /support

/offices

Millipore, Ultracel, Immobilon, and Amicon are registered trademarks of Millipore Corporation.

The M mark, Advancing Life Science Together, PureProteome, Magna GrIP, SNAP i.d., and

Spray & Glow are trademarks of Millipore Corporation.

Lit. No. DS1060EN00 11/08 BP-GEN-08-01172

Printed in the U.S.A.

© 2008 Millipore Corporation, Billerica, MA 01821 U.S.A. All rights reserved.