Anti-PDPK1 (phospho S393) antibody [EPR19818] (ab206340)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19818] to PDPK1 (phospho S393)
- Suitable for: WB, IP, Dot blot
- Reacts with: Human, Recombinant fragment
Overview
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Product name
Anti-PDPK1 (phospho S393) antibody [EPR19818]
See all PDPK1 primary antibodies -
Description
Rabbit monoclonal [EPR19818] to PDPK1 (phospho S393) -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Dot Recombinant fragmentIP HumanWB Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK-293T whole cell lysate transfected with PDPK1 (WT). IP: HEK-293T whole cell lysate transfected with PDPK1 (WT).
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19818 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-PDPK1 (phospho S393) antibody [EPR19818] (ab206340) at 1/5000 dilution
Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate transfected with PDPK1 S393A mutant
Lane 2 : HEK-293T whole cell lysate transfected with PDPK1 (WT)
Lane 3 : HEK-293T whole cell lysate transfected with PDPK1 (WT), then treated with alkaline phosphatase for 1 hour
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 63 kDa
Observed band size: 60,63 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
These plasmids were kindly provided by our collaborator Dr. Yi Rao, Peking University.
Based on sequence alignment, the antigen can recognize 5 isoforms, the predicted MW are 63kDa, 58 kDa, 60 kDa, 48 kDa and 51kDa, respectively.
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Dot blot analysis of PDPK1 (phospho S393) labeled with ab206340 at 1/1000 dilution.
Lane 1: PDPK1 (phospho S393) peptide(aa391-401)
Lane 2: PDPK1 (phospho S393) peptide(aa386-396)
Lane 3: PDPK1 non-phospho peptide(aa386-401)
Lane 4: PDPK1 (phospho S396) peptide (aa392-401)
Lane 5: PDPK1 (phospho S396) peptide(aa393-402)
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated (ab97051) at 1/20000 dilution was used as secondary antibody.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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All lanes : Anti-PDPK1 (phospho S393) antibody [EPR19818] (ab206340) at 1/1000 dilution
Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate transfected with PDPK1 WT
Lane 2 : HEK-293T whole cell lysate transfected with PDPK1 (WT), then treated with alkaline phosphatase for 1 hour
Lane 3 : HEK-293T whole cell lysate transfected with PDPK1 S393A mutant
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/2000 dilution
Predicted band size: 63 kDa
Observed band size: 60,63 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsBlocking/Dilution buffer: 5% BSA/TBST.
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PDPK1 (phospho S393) was immunoprecipitated from 0.35 mg of HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate, transfected with PDPK1 (WT), with ab206340 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab206340 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HEK-293T whole cell lysate transfected with PDPK1 (WT), 10 μg (Input).
Lane 2: ab206340 IP in HEK-293T whole cell lysate transfected with PDPK1 (WT).
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab206340 in HEK-293T whole cell lysate transfected with PDPK1 (WT).
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
The plasmids were kindly provided by our collaborator Dr. Yi Rao, Peking University.
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