Protein A Sepharose® (ab193256)
Key features and details
- Sample type: Ascites Fluid, Cell culture media, Cell culture supernatant, Serum
Overview
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Product name
Protein A Sepharose® -
Sample type
Cell culture supernatant, Serum, Cell culture media, Ascites Fluid -
Product overview
Contents:
Supplied as a 50% slurry in 20% Ethanol.
Features:
High binding capacity = Binding of IgG ≥16 mg human or rabbit IgG/mL Protein A-Sepharose®.
Minimal leaching of the ligand
Flow Rate Tested* = 2.07 mL/min.
*Test condition: = Calculations based on the time required to pass 18 mL of water through 2 mL settled beads (column diameter 1.5 cm).
Usage = Reusable for up to 10 times without significant loss of binding capacity.
Store beads at 4°C.
The beads may be damaged above 40°C.
DO NOT FREEZE.
Wash beads 3 times with 3x bead volume of desired buffer before use.
Applications:
- Purification of monoclonal and polyclonal antibodies from culture media, serum, ascites fluid or hybridoma supernatants.
- Isolation of antibody/antigen complexes in immunoprecipitation experiments, since only the Fc region is involved in antibody binding and the Fab region is available for binding antigen.
Sepharose is a registered trademark of GE Healthcare
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Notes
This product is manufactured by BioVision, an Abcam company and was previously called 6501 Protein A Sepharose. 6501-5 is the same size as the 5 ml size of ab193256.
Protein A Sepharose® beads are prepared by covalently coupling recombinant Protein A to 6% cross-linked Sepharose® beads. Protein A is a genetically engineered protein containing five IgG-binding regions of native Protein A. The cell wall binding region, albumin binding region and other non-specific regions have been eliminated from the recombinant Protein A to ensure maximum specific IgG binding. The coupling technique is optimized to give a higher binding capacity for IgG and minimum leaching of recombinant Protein A. The IgG binding capacity of Protein A Sepharose® is ≥ 16 mg human or rabbit IgG per mL of wet beads. Protein A Sepharose® beads display high chemical and physical stability as well as high flow rate, hydrophilicity and high gel strength. This product can be used for IgG purification and immunoprecipitation.
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Tested applications
Suitable for: IP, Purificationmore details
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 ml 5 ml 25 ml 100 ml Protein A Sepharose® 1 x 1ml 1 x 5ml 1 x 25ml 1 x 100ml -
Research areas
Associated products
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab193256 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| IP |
Use at an assay dependent concentration.
Isolation of antibody/antigen complexes in immunoprecipitation experiments, since only the Fc region is involved in antibody binding and the Fab region is available for binding antigen. |
|
| Purification |
Use at an assay dependent concentration.
Purification of monoclonal and polyclonal antibodies from culture media, serum, ascites fluid or hybridoma supernatants. |
| Notes |
|---|
|
IP
Use at an assay dependent concentration. Isolation of antibody/antigen complexes in immunoprecipitation experiments, since only the Fc region is involved in antibody binding and the Fab region is available for binding antigen. |
|
Purification
Use at an assay dependent concentration. Purification of monoclonal and polyclonal antibodies from culture media, serum, ascites fluid or hybridoma supernatants. |
Images
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Immunoprecipitation - Protein A Sepharose® (ab193256)Image from Arnold et al., J Exp Clin Cancer Res., 39(1):205; doi: 10.1186/s13046-020-01712-w. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
Overexpression of TTLL4 in MDA-MB231 cells increases MT-glutamylation.b Polyglutamylated proteins were immunoprecipitated from control and TTLL4 plus cells, using an antibody against polyglutamylation modification (GT335). Cell lysates ("input"), supernatants ("S/N") from cell lysates incubated with GT335-coupled beads and proteins bound to GT335 coupled beads ("beads") were analyzed
by Western blotting for β-tubulin and NAP-1 levels. IgG signals served as loading control. The lower band appearing in the NAP-1 blot is a residue signal of ?-tubulin antibody because the same membrane was used to probe against all 3 proteins. -
Immunoprecipitation - Protein A Sepharose® (ab193256)Image from O'Leary et al., Sci Rep., 8(1):15360; doi: 10.1038/s41598-018-33840-x. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Immunoprecipitation of resistin from obese subcutaneous adipose conditioned medium secretome (OB ACM) improves myogenesis. Resistin was immunoprecipitated from OB ACM using resistin antibody-agarose bead conjugates (OB ACM - resistin IP). IgG isotype antibody control-agarose bead conjugates were used on the same samples as a control (OB ACM).(A) Resistin protein is detected by immunoblotting of resistin-antibody lysates but not IgG control lysates following immunopreciptation.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (7)
ab193256 has been referenced in 7 publications.
- Zhou C et al. FGF8 and BMP2 mediated dynamic regulation of dental mesenchyme proliferation and differentiation via Lhx8/Suv39h1 complex. J Cell Mol Med 25:3051-3062 (2021). PubMed: 33580754
- Arnold J et al. Tubulin Tyrosine Ligase Like 4 (TTLL4) overexpression in breast cancer cells is associated with brain metastasis and alters exosome biogenesis. J Exp Clin Cancer Res 39:205 (2020). PubMed: 32998758
- Prasad V et al. The UPR sensor IRE1a and the adenovirus E3-19K glycoprotein sustain persistent and lytic infections. Nat Commun 11:1997 (2020). PubMed: 32332742
- Marenne G et al. Exome Sequencing Identifies Genes and Gene Sets Contributing to Severe Childhood Obesity, Linking PHIP Variants to Repressed POMC Transcription. Cell Metab 31:1107-1119.e12 (2020). PubMed: 32492392
- Das S et al. CXCR7: A key neuroprotective molecule against alarmin HMGB1 mediated CNS pathophysiology and subsequent memory impairment. Brain Behav Immun 82:319-337 (2019). PubMed: 31505255
- O'Leary MF et al. Obese subcutaneous adipose tissue impairs human myogenesis, particularly in old skeletal muscle, via resistin-mediated activation of NF?B. Sci Rep 8:15360 (2018). PubMed: 30337633
- Liu YG et al. Abrogation of cathepsin C by Helicobacter pylori impairs neutrophil activation to promote gastric infection. FASEB J N/A:fj201802016RR (2018). PubMed: 30596522