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Purified anti-PDPK1 Phospho (Ser241) Antibody

Pricing & Availability
Clone
A19011A (See other available formats)
Regulatory Status
RUO
Other Names
PDK1, PkB Kinase, 3-Phosphoinositide Dependent Protein Kinase 1
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
1_A19011A_PURE_PDPK1_Phospho_Antibody_2_102920
Whole cell extracts (15 µg total protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:500 dilution) of purified anti-PDKP1 Phospho (Ser241) (clone A19011A) for 2 hours at room temperature. To confirm phospho-specificity of clone A19011A, a duplicate membrane was pre-treated with lambda protein phosphatase prior to incubation with the antibody. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG (Cat. No. 405306) at a 1:3000 dilution. Total PDPK1 levels were confirmed by probing membranes with purified anti- PDPK1 antibody (clone W17215A) (Cat. No. 617602) at 1.0 µg/mL (1:500 dilution). Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker.
  • 1_A19011A_PURE_PDPK1_Phospho_Antibody_2_102920
    Whole cell extracts (15 µg total protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:500 dilution) of purified anti-PDKP1 Phospho (Ser241) (clone A19011A) for 2 hours at room temperature. To confirm phospho-specificity of clone A19011A, a duplicate membrane was pre-treated with lambda protein phosphatase prior to incubation with the antibody. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG (Cat. No. 405306) at a 1:3000 dilution. Total PDPK1 levels were confirmed by probing membranes with purified anti- PDPK1 antibody (clone W17215A) (Cat. No. 617602) at 1.0 µg/mL (1:500 dilution). Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker.
  • 2_A19011A_PURE_PDPK1_Phospho_Antibody_102920
    Whole cell extracts (15 µg total protein) from HeLa cells transfected with non-targeting control siRNA (siCON) or siRNA targeting PDPK1 (siPDPK1) resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:500 dilution) purified anti-PDKP1 Phospho (Ser241) (clone A19011A) for 2 hours at room temperature. To confirm phospho-specificity of clone A19011A, a duplicate membrane was pre-treated with lambda protein phosphatase prior to incubation with the antibody. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG (Cat. No. 405306) at a 1:3000 dilution. Total PDPK1 levels were confirmed by probing membranes with purified anti-PDPK1 antibody (clone W17215A) (Cat. No. 617602) at 1.0 µg/mL (1:500 dilution). Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker.
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939601 25 µg £97
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939602 100 µg £226
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Description

3-Phosphoinositide-Dependent Protein Kinase 1 (PDPK1) is a serine/threonine kinase that plays a central role in diverse cellular pathways, and can function as a critical activator of several prosurvival and oncogenic protein kinases. PDPK1 functions by phosphorylating and activating members of the AGC family of serine kinases, including Akt1, protein kinases A and C, and p70 ribosomal protein S6 kinase. Loss or reduction of PDPK1 in the thymus results in defective T-cell differentiation and proliferative expansion.

Serine 241 of PDPK1 is an autophosphorylation site located within the activation loop of the protein and is required for full enzymatic function. This modification is considered a candidate pharmacodynamic biomarker of efficacy for PDPK1 inhibitors

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Synthetic peptide corresponding to human PDPK1 phosphorylated at serine 241
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
WB-KO/KD - Validated
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 1.0 µg/mL. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

-A19011A was tested for ICC using 4% PFA-fixed HeLa cells permeabilized with either methanol or Triton X-100. While staining was observed, the signal was PDPK1-independent. We therefore do not recommend the clone for this application.

-A19011A is predicted to recognize the residue corresponding to Ser241 in multiple PDPK1 isoforms due to the presence of the immunizing sequence in each isoform.

-A19011A is predicted to recognize rat PDPK1 phosphorylated at Ser244 due to complete sequence homology between the immunizing peptide and the rat PDPK1 ortholog.

-A19011A may cross-react with PSK1-α when phosphorylated at Ser181 due to partial sequence homology with the immunizing peptide

RRID
AB_2888741 (BioLegend Cat. No. 939601)
AB_2888741 (BioLegend Cat. No. 939602)

Antigen Details

Structure
PDPK1 is a 556 amino acid protein with a predicted molecular weight of 63 kD. Four other PDPK1 isoforms have been described, with predicted molecular weights of 48, 51, 58, and 60 kD.
Distribution

Ubiquitously expressed/cytosolic

Function
Serine-threonine protein kinase/cell signaling
Biology Area
Cell Biology
Molecular Family
Protein Kinases/Phosphatase
Antigen References
1. Le Good JA, et al. 1998. Science. 281:2042.
2. Casamayor A, et al. 1999. Biochem J. 342 Pt2):287.
3. Dong LQ, et al. 2000. Proc. Natl. Acad. Sci. 97:5089.
4. Hinton HJ, et al. 2004. Nat. Immunol. 5:539.
5. Nagasima K, et al. 2005. J. Biol. Chem. 286:6433
Gene ID
5170 View all products for this Gene ID
UniProt
View information about PDPK on UniProt.org
Go To Top Version: 1    Revision Date: 10/29/2020

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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