Recombinant
RabMAb

Recombinant Anti-SIK1 (phospho T182) + SIK2 (phospho T175) + SIK3 (phospho T163) antibody [EPR19196] (ab199474)

Overview

  • Product name

    Anti-SIK1 (phospho T182) + SIK2 (phospho T175) + SIK3 (phospho T163) antibody [EPR19196]
  • Description

    Rabbit monoclonal [EPR19196] to SIK1 (phospho T182) + SIK2 (phospho T175) + SIK3 (phospho T163)
  • Host species

    Rabbit
  • Specificity

    Expected to react with Snf1lk (when phosphorylated at T182) and SIK3 ((when phosphorylated at T163).
  • Tested applications

    Suitable for: Dot blot, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human SIK2 aa 150-250. The exact sequence is proprietary. aa177 modified from C to S
    Database link: Q9H0K1

  • Positive control

    • WB: HEK-293T transfected with human Snf1lk or Snf1lk2 or SIK3 (WT) whole cell lysate.
  • General notes

     This product was previously labelled as Snf1lk2, Snf1lk+Snf1lk2+SIK3

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab199474 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
Dot blot 1/1000.
WB 1/1000. Detects a band of approximately 85, 104, 140 kDa.

Target

Images

  • All lanes : Anti-SIK1 (phospho T182) + SIK2 (phospho T175) + SIK3 (phospho T163) antibody [EPR19196] (ab199474) at 1/1000 dilution

    Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with empty vector (control), whole cell lysate
    Lane 2 : HEK-293T transfected with a human SIK3 (WT) expression vector, whole cell lysate
    Lane 3 : HEK-293T transfected with a human SIK3 (WT) expression vector, then treated with alkaline phosphatase for 1 hour, whole cell lysate
    Lane 4 : HEK-293T transfected with a human SIK3 T163A mutant expression vector, whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Observed band size: 140 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

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

  • All lanes : Anti-SIK1 (phospho T182) + SIK2 (phospho T175) + SIK3 (phospho T163) antibody [EPR19196] (ab199474) at 1/1000 dilution

    Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with an empty vector (control), whole cell lysate
    Lane 2 : HEK-293T transfected with a human SIK2 (WT) expression vector, whole cell lysate
    Lane 3 : HEK-293T transfected with a human SIK2 (WT) expression vector, then treated with alkaline phosphatase for 1 hour, whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Observed band size: 104 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

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

  • All lanes : Anti-SIK1 (phospho T182) + SIK2 (phospho T175) + SIK3 (phospho T163) antibody [EPR19196] (ab199474) at 1/5000 dilution

    Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with an empty vector (control), whole cell lysate
    Lane 2 : HEK-293T transfected with a human SIK1 (WT) expression vector, whole cell lysate
    Lane 3 : HEK-293T transfected with a human SIK1 (WT) expression vector, then treated with alkaline phosphatase for 1 hour, whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Observed band size: 85 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

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

  • Dot blot analysis of Snf1lk2 (phospho T175) labeled with ab199474 at 1/1,000 dilution.

    Lane 1: Snf1lk2 (phospho T175) peptide

    Lane 2: Snf1lk2 non-phospho peptide

    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100,000 dilution was used as secondary antibody.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

References

ab199474 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (A549 cells (Wild Type and LKB1 addback))
Gel Running Conditions
Reduced Denaturing (8%)
Loading amount
30 µg
Specification
A549 cells (Wild Type and LKB1 addback)
Blocking step
Milk as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 05 2019

Application
Western blot
Sample
Mouse Cell lysate - whole cell (HEK 293T)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
20 µg
Specification
HEK 293T
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. liu yuxiang

Verified customer

Submitted Nov 10 2017

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