| Distributors | Register Now  |  
Shopping cart( 0 Items )Order Status  
 
Home
emailprintBook mark

Protein A Separopore® 4B Cartridge

 

MSRP: $321.21*

Price: $274.54*

Add to Cart: QTY:



Download COA

Protein A Separopore® 4B

Affinity matrix with immobilized Protein-A (from staphylococcus aureus) by CNBr activation coupling method. For purification and/or fractionation of IgG subclasses. Reliable tool to purify monoclonal and polyclonal IgG from ascies, serum and cell culture supernatants.

Provided as pre-packed, ready-to-use Protein A Separopore® Chromatography Cartridges. Conveniently packed in a 2.5 ml in-line column capable of running at low pressures. Fittings to attach with any HPLC or FPLC systems are provided separately.

Note: Separopore® is a cost-effective equivalent to Sepharose® in all of its physical properties and binding characteristics.

IP with Protein A

• Add 3 ml ice cold RIPA buffer to cell monolayer (or approximately 2–5 x 107 suspension cells in flask) and incubate at 4° C for 10 minutes. For suspension cells, add the RIPA buffer to washed cell pellet in a 15 ml conical centrifuge tube.
• Disrupt cells by repeated aspiration through a 21 gauge needle and transfer to a 15 ml conical centrifuge tube.
• Wash cell culture plate with additional 1.0 ml ice cold RIPA buffer and combine with original extract.
• Pellet cellular debris by centrifugation at 10,000xg for 10 minutes at 4° C. Transfer supernatant to a 15 ml conical centrifuge tube kept on ice. Preclear lysate (optional step) by adding 1.0 μg of the appropriate control IgG (normal mouse, rat, rabbit or goat IgG, corresponding to the host species of the primary antibody)towns add 20 μl of resuspended volume of Protein A-Separopore. Incubate at 4° C for 30 minutes.
• Pellet beads by centrifugation at 2,500 rpm (approximately 1,000xg) for 5 minutes at 4° C. Transfer supernatant to a fresh 15 ml conical centrifuge tube kept on ice.
• Transfer 1 ml of the above cell lysate, or approximately 100–500 μg total cellular protein, to a 1.5 ml microcentrifuge tube. Add 1–10 μl (i.e., 0.2–2 μg) primary antibody (optimal antibody concentration should be determined by titration) and incubate for 1 hour at 4° C. longer incubation. May be. Required for some low titter antibodies.
• Add 20 μl of resuspended volume of Protein A-Separopore.  Cap tubes and incubate at 4° C on a rocker platform or rotating device for 1 hour.
• Collect immunoprecipitates by centrifugation at 2,500 rpm (approximately 1,000xg) for 5 minutes at 4° C. Carefully aspirate and discard supernatant.
• Wash pellet 4 times with 1.0 ml RIPA buffer (more stringent) or PBS (less stringent), each time repeating centrifugation step above.
• After final wash, aspirate and discard supernatant and resuspend pellet in 40 μl of 1x electrophoresis sample buffer.
• Boil samples for 2minutes and analyze 20 μl aliquots by SDS-PAGE and follow it by Western blot analysis or auto radiography if sample was labeled.


Catalog No.:20170020-1
Size:5 mL
Brand Name:bioPLUS™, Separopore®
Hazmat Shipping:Non-hazardous
Storage:2-8°C
Matrix:Cross-linked agarose beads, 4%.
Particle Size Range:45-165μm
Binding Capacity:approximately 20mg human IgG/ml drained gel.
Suspension:Supplied as suspension (50% slurry) in 0.5M NaCl
pH Stability:3-9 (for use), 2-10 (for cleaning)
Ligand Coupling Method:CNBr activation
Ligand:Native protein A from Staphylococcus aureus.

*Cold Shipping and packing charges apply.

  Protocols

Please check the protocols on www.biolabprotocols.com

If you have any protocols or usage recommendations on any of our products that to share with other scientists. Please feel free to submit them to us at info@biolabprotocols.com and we will be delighted to post them for you on www.biolabprotocols.com


  References and Citations

Share your publication or characterization data with us.
Please contact us to post reference.
E-mail : Send email


  Customer also viewed the following products

Co-IMAC 5 x 2mL Columns Co-IMAC 5 x 2mL Columns

Co-immobilized metal affinity chromatography (IMAC) columns consist of Co-Chelated Separopore®matrix conveniently prepacked spin columns for gravity flow chromatography. Co-IMAC Columns separ... more

Aprotinin Separopore® 6B-CL OnePass™ Column Aprotinin Separopore® 6B-CL OnePass™ Column

bioWORLD's immonibilzed aprotinin is pure aprotinin coupled to Separopore® (Agarose) 6B-CL. It is designed to remove serine proteases from cell or tissue lysates. Aprotinin, a monomeric protein in... more

Protein A Separopore® 4B OnePass™ Column Protein A Separopore® 4B OnePass™ Column

Protein A Separopore® 4B Affinity matrix with immobilized Protein-A (from staphylococcus aureus) by CNBr activation coupling method. For purification and/or fractionation of IgG subclasses. Relia... more

Fetuin Separopore® 4B OnePass™ Column Fetuin Separopore® 4B OnePass™ Column

Bovine fetuin has N-linked glycans with terminal Siaα2-3Galβ1-4GlcNAc, Siaα2-6Galβ1-4GlcNAc and unsubstituted Galβ1-4GlcNAc residues. It also has O-linked glycans that inclu... more

Ask a question

 
MagneZoom™ ParaMagnetic Bead Station
Featured Items: