Antibodies, Proteins, Kits and Reagents for Life Science

298 185 ₸ Product size

100 µl 298 185 ₸

Order now and get it on Wednesday March 17, 2021

Anti-PDPK1 (phospho S396) antibody [EPR19819] (ab206341)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19819] to PDPK1 (phospho S396)
Suitable for: IP, WB, Dot blot
Reacts with: Human, Recombinant fragment

Product name

Anti-PDPK1 (phospho S396) antibody [EPR19819]
See all PDPK1 primary antibodies
Description

Rabbit monoclonal [EPR19819] to PDPK1 (phospho S396)
Host species

Rabbit

Tested Applications & Species

Application	Species
Dot	Recombinant fragment
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HEK-293T whole cell lysate transfected with PDPK1 (WT). IP: HEK-293T transfected with PDPK1 whole cell lysate.
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
For more information see here.
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.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19819
Isotype

IgG
Research areas

Western blot - Anti-PDPK1 (phospho S396) antibody [EPR19819] (ab206341)

All lanes : Anti-PDPK1 (phospho S396) antibody [EPR19819] (ab206341) 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 S396A 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 minutes

5% NFDM/TBST: Blocking and dilution buffer.

The plasmids were kindly provided by our collaborator Dr. Yi Rao, Peking University.

Based on sequence alignment, this antigen can recognize 5 isoforms, the predicted MW are 63kDa, 58 kDa, 60 kDa, 48 kDa and 51kDa, respectively.
Dot Blot - Anti-PDPK1 (phospho S396) antibody [EPR19819] (ab206341)

Dot blot analysis of PDPK1 (phospho S396) labeled with ab206341 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 (aa392-402)

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.
Western blot - Anti-PDPK1 (phospho S396) antibody [EPR19819] (ab206341) This data was kindly provided by Dr. Yi Rao, Peking University.

All lanes : Anti-PDPK1 (phospho S396) antibody [EPR19819] (ab206341) 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 S396A 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 seconds

Blocking/Dilution buffer: 5% BSA/TBST.
Immunoprecipitation - Anti-PDPK1 (phospho S396) antibody [EPR19819] (ab206341)

PDPK1 (phospho S396) 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, with ab206341 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab206341 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: HEK-293T transfected with PDPK1 whole cell lysate, 10 μg (Input).

Lane 2: ab206341 IP in HEK-293T transfected with PDPK1 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab206341 in HEK-293T transfected with PDPK1 whole cell lysate.

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.
Anti-PDPK1 (phospho S396) antibody [EPR19819] (ab206341)

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