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  • Lysates were prepared from the indicated yeast deletion strains expressing HA-tagged PP2A C subunit (designated PPH21 in yeast) and immunoprecipitated using an anti HA-tag antibody. The ppe1D strain lacks PP2A methylesterase, and thus accumulates (hyper)methylated PPH21. The ppm1D strain lacks PP2A methyltransferase and does not contain any methylated PPH21. Samples were separated by 10% SDS-PAGE, blotted onto nitrocellulose and probed with the 2A10 antibody with ECL visualisation.
  • PP2A alpha (methyl L309) Monoclonal Antibody [2A10]

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PP2A alpha (methyl L309) Monoclonal Antibody [2A10]

Catalogue Number: IQ556

Catalogue Number: IQ556HRP

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  • £328.90

Datasheet

Download Variant Datasheet
Primary Description: Mouse Anti-PP2A alpha (methyl L309) [2A10]
Primary Description: Mouse Anti-PP2A alpha (methyl L309) [2A10]-
Target Antigen: PP2A alpha (methyl L309)
Target Antigen: PP2A alpha (methyl L309)
Catalogue Number: IQ556
Catalogue Number: IQ556HRP
Quantity: 0.1mg
Quantity: 0.05mg
Concentration: 1mg/ml
Concentration: 1mg/ml
Clone: 2A10
Clone: 2A10
Host: Mouse
Host: Mouse
Isotype: IgG1
Isotype: IgG1
Myeloma/Fusion Partners: X63-Ag8.653
Myeloma/Fusion Partners: X63-Ag8.653
Immunogen: A KLH-coupled peptide with the sequence: KLH-C-ßA-RRTPDYFL-OMe. The c-terminal sequence (as well as the carboxy-methylation) is absolutely conserved from human, mouse to yeast
Immunogen: A KLH-coupled peptide with the sequence: KLH-C-ßA-RRTPDYFL-OMe. The c-terminal sequence (as well as the carboxy-methylation) is absolutely conserved from human, mouse to yeast
Species Reactivity: Mouse, Rat, Human.Predicted to work with: Saccharomyces cerevisiae
Species Reactivity: Mouse, Rat, Human.Predicted to work with: Saccharomyces cerevisiae
Purification: Protein G
Purification: Protein G
Format: Purified antibody (from supernatant) containing PBS + 0.1% sodium azide
Format: Purified IgG conjugated to Horseradish Peroxidase using Innova Biosciences Lightning-Link®, supplied in Phosphate buffered saline (PBS) containing 0.01% Thiomersal
Applications: WB, IP
Applications: WB, IP
Dilutions: Optimal antibody dilution should be determined by titration, however as a guideline try; WB: 1;500. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa)
Dilutions: Optimal antibody dilution should be determined by titration, however as a guideline try; WB: 1;500. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa)
Also Known As: PP2A alpha antibody;PP2A C antibody;PP2A-alpha antibody;PP2AA_HUMAN antibody;PP2Ac antibody;PP2CA antibody;PPP2CA antibody;Protein phosphatase 2, catalytic subunit, alpha isoform antibody;Replication protein C antibody
Also Known As: PP2A alpha antibody;PP2A C antibody;PP2A-alpha antibody;PP2AA_HUMAN antibody;PP2Ac antibody;PP2CA antibody;PPP2CA antibody;Protein phosphatase 2, catalytic subunit, alpha isoform antibody;Replication protein C antibody
Pubmed ID(s): 22610379, 21698000, 20636478, 17550305, 15535135, 15099019, 12952889
Pubmed ID(s): 22610379, 21698000, 20636478, 17550305, 15535135, 15099019, 12952889
Entrez Gene ID(s): 5515, 19052, 24672
Entrez Gene ID(s): 5515, 19052, 24672
SwissProt ID(s): P67775, P63330, P63331

Citations

Citation Count: 7
Citations:

Benziane B  et al. Activation of AMP-activated Protein Kinase Stimulates Na+,K+-ATPase Activity in Skeletal Muscle Cells. J Biol Chem 287:23451-63 (2012). WB
View Source


Li T  et al. Microcystin-LR (MCLR) Induces a Compensation of PP2A Activity Mediated by a4 Protein in HEK293 Cells. Int J Biol Sci 7:740-52 (2011). WB ; Human
View Source


Finnegan S  et al. A stress survival response in retinal cells mediated through inhibition of the serine / threonine phosphatase PP2A. Eur J Neurosci : (2010). WB ; Mouse
View Source


Hombauer H  et al. Generation of active protein phosphatase 2A is coupled to holoenzyme assembly. PLoS Biol 5:e155 (2007)
View Source


Sontag E  et al. Downregulation of protein phosphatase 2A carboxyl methylation and methyltransferase may contribute to Alzheimer disease pathogenesis. J Neuropathol Exp Neurol 63:1080-91 (2004)
View Source


Sontag E  et al. Altered expression levels of the protein phosphatase 2A ABalphaC enzyme are associated with Alzheimer disease pathology. J Neuropathol Exp Neurol 63:287-301 (2004)
View Source


Fellner T  et al. A novel and essential mechanism determining specificity and activity of protein phosphatase 2A (PP2A) in vivo. Genes Dev 17:2138-50 (2003)
View Source


Protocols

Immunofluorescence protocol - Formaldehyde fixation
  1. Collect cells from T.c.unit and remove media from petri dish using suction.
  2. Wash with 1x PBS and remove.
  3. Incubate cells in pre-warm (37°C) Para-Formaldehyde for 12 minutes at room temperature on an orbital shaker.
  4. Remove PFA and incubate in 0.5% Triton X-IOO in 1x PBS for 5 minutes at room temperature.
  5. Prepare blocking reagent, this is also the antibody diluent.
  6. Wash cells 2x with 1x PBS at room temperature, for 4 minutes/wash on an orbital shaker.
  7. Block with 1 % NCS and 1x PBS for 30 minutes at room temperature.
  8. Prepare primary antibodies (50?l/coverslip) and moist staining chambers.
  9. Wash cells 2x with lx PBS at room temperature and air dry briefly.
  10. Incubate with primary antibody for 1 hr at room temperature in the dark in staining chambers. During this time prepare the secondary antibody.
  11. Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)
  12. Incubate with secondary antibody for 1 hour at room temperature in the dark in staining chambers.
  13. Wash cells 5x with 1x PBS.
  14. Mount in Dapi.

Solutions (prepare fresh the same day of staining).

  • * 1x Phosphate buffered saline.
  • * Blocking reagent: 1% NCS in 1x PBS (use fresh l0x PBS).
  • * Fixation solution: 3.5% Para formaldehyde.

1.75g PFA in 20 ml d.H20 plus 5 drops 1M NaOH. Stir on a hot plate at 50-60°C until dissolved. Add 4 drops IN HCI and check pH indicator strip. PH should be 7.4. Complete volume with d.H20 to 25ml and add 25ml 2xPBS. Check pH before adding to cover slips.

Immunofluorescence protocol - Methanol/acetone fixation
  1. Collect cells from T.C.unit and remove media from petri dish using suction.
  2. Wash with 1x PBS and remove.
  3. Fix cells with cold methanol: acetone 1: 1 for 10 minutes on ice.
  4. Prepare blocking reagent, this is also the diluent for the antibodies.
  5. Remove fixative and wash cells 3x with Ix PBS at RT, for 4 minutes/wash on orbital shaker.
  6. Block with 1% NCS and Ix PBS for 30 minutes at RT.
  7. Prepare primary antibodies (50?l/coverslip) and moist staining chambers.
  8. Wash cells 2x with 1 x PBS at RT and air dry for approximately 7 minutes.
  9. Incubate with primary antibody for 1 hr at RT in the dark in staining chambers. During this time prepare secondary antibody.
  10. Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)
  11. Incubate with secondary antibody for 1 hr at R T in the dark in staining chambers.
  12. Wash cells 5x with 1x PBS.
  13. Mount in Dapi.

Solutions (prepare fresh the same day of staining)

  • * 1x Phosphate buffered saline.
  • * Blocking reagent: 1% NCS in 1x PBS (use fresh 10x PBS).
  • * Fixation solution: methanol:acetone 1: 1 ice cold.
Western Blotting Protocol
  1. Transfer gel to PDVF or nitrocellulose membrane
  2. Place membrane in plastic tray in blocking buffer for one hour with agitation
  3. Rinse in wash buffer
  4. Incubate in wash buffer plus primary antibody for one hour
  5. Wash 6 X 5 minutes with wash buffer
  6. Incubate in wash buffer plus secondary antibody for one hour
  7. Wash 6X 5 minutes with wash buffer
  8. Detect (e.g. ECL, Amersham according to manufacturers instructions)
Wash buffer

PBS + 0.1% Tween 20

Blocking buffer

Wash buffer + 5% dried milk powder

The concentration of antibodies used depends on each antibody, the amount of antigen and the detection method used. Generally, dilution is in the range of a few hundred times dilution to a few thousand times dilution, but usually has to be determined empirically.

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We are strongly committed to providing the best quality products, and the best service possible.

Should any product not perform as described in the product literature, it will be replaced or a full refund will be given. The customer must notify ImmuQuest within 30 days after the goods have been received to request a replacement, forwarding complete test data as requested by ImmuQuest.

If items are ordered incorrectly by the customer, ImmuQuest will consider taking them back as long as they have been stored correctly and have not been opened or tampered with. Such orders may be subject to a 15% restocking charge on the items plus any shipping costs.

Requests for returns must have prior authorization from ImmuQuest, and must be made within 7 days of receipt of the items. Items must be returned in the same or equivalent packaging as originally dispatched, and by an equivalent method of delivery.

Product Guarantee

We are strongly committed to providing the best quality products, and the best service possible. In order to do this we depend on your feedback.

Should any product not perform as described in the product literature, it will be replaced or a full refund will be given. The customer must notify ImmuQuest within 30 days after the goods have been received to request a replacement, forwarding complete test data as requested by ImmuQuest.

If items are ordered incorrectly by the customer, ImmuQuest will consider taking them back as long as they have been stored correctly and have not been opened or tampered with. Such orders may be subject to a 15% restocking charge on the items plus any shipping costs.

Requests for returns must have prior authorization from ImmuQuest, and must be made within 7 days of receipt of the items. Items must be returned in the same or equivalent packaging as originally dispatched, and by an equivalent method of delivery.

If you have any questions please contact us using the following details:

ImmuQuest Ltd
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Stokesley Business Park
24 Ellerbeck Way
Stokesley
TS9 5JZ
UK

Tel: 01642 713533
Fax: 01642 713988
International Tel: +44 1642 713533
International Fax: +44 1642 713988

When placing an order ImmuQuest require a purchase order number, plus name and contact details of the purchaser, and the person who will be using the product. ImmuQuest will also need a VAT number for customers in the European Union.UK customers that are VAT exempt need to fax an exemption certificate.

Orders can be placed either by our website, via email or by mail. Please see our contact us page for more details. All orders are subject to availability. Prices of products do not include shipping, VAT or import duties where these are applicable.

Price and other information provided are subject to change without notice. While every effort is made to keep information provided Up to date, ImmuQuest will not be liable if errors should occur in such information. Formal acceptance of an order will take place when the goods are dispatched. If prices should be changed between the time of receipt of an order and dispatch, ImmuQuest will contact you in advance.