Overview

  • Product name

  • Description

    Rabbit polyclonal to SGK1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Rabbit, Chicken
  • Immunogen

    Synthetic peptide corresponding to Human SGK1 aa 300-400 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab43605)

  • Positive control

    • WB: HeLa, Jurkat, A431, BT549, T47D and NIH 3T3 whole cell lysates and human pancreas, lung and brain tissue lysates. ICC/IF: HeLa cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab43606 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 49 kDa (predicted molecular weight: 49 kDa).
ICC/IF Use a concentration of 10 µg/ml.
IHC-P 1/2000.

Target

  • Function

    Protein kinase that plays an important role in cellular stress response. Activates certain potassium, sodium, and chloride channels, suggesting an involvement in the regulation of processes such as cell survival, neuronal excitability and renal sodium excretion. Sustained high levels and activity may contribute to conditions such as hypertension and diabetic nephropathy. Mediates cell survival signals, phosphorylates and negatively regulates pro-apoptotic FOXO3A. Phosphorylates NEDD4L, which leads to its inactivation and to the subsequent activation of various channels and transporters such as ENaC, KCNA3/Kv1.3 or EAAT1. Isoform 2 exhibited a greater effect on cell plasma membrane expression of ENaC and Na(+) transport than isoform 1.
  • Tissue specificity

    Expressed in most tissues with highest levels in the pancreas, followed by placenta, kidney and lung. Isoform 2 is strongly expressed in brain and pancreas, weaker in heart, placenta, lung, liver and skeletal muscle.
  • Sequence similarities

    Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family.
    Contains 1 AGC-kinase C-terminal domain.
    Contains 1 protein kinase domain.
  • Domain

    Isoform 2 subcellular localization at the plasma membrane is mediated by the sequences within the first 120 amino acids.
  • Post-translational
    modifications

    Regulated by phosphorylation. Phosphoinositide 3-kinase (PI3-kinase) pathway promotes phosphorylation at Ser-422 which in turn increases the phosphorylation of Thr-256 by PDPK1.
    Ubiquitinated by NEDD4L; which promotes proteasomal degradation. Ubiquitinated by SYVN1 at the endoplasmic reticulum; which promotes rapid proteasomal degradation and maintains a high turnover rate in resting cells. Isoform 2 shows enhanced stability. Isoform 2 resistance to proteasomal degradation is mediated by the sequences within the first 120-amino acid.
  • Cellular localization

    Cell membrane and Cytoplasm. Nucleus. Endoplasmic reticulum. Nuclear, upon phosphorylation.
  • Information by UniProt
  • Database links

  • Alternative names

    • OTTHUMP00000017247 antibody
    • Serine/threonine protein kinase SGK antibody
    • Serine/threonine protein kinase Sgk1 antibody
    • Serine/threonine-protein kinase Sgk1 antibody
    • Serum and glucocorticoid regulated kinase antibody
    • Serum/glucocorticoid regulated kinase 1 antibody
    • Serum/glucocorticoid regulated kinase antibody
    • Serum/glucocorticoid-regulated kinase 1 antibody
    • SGK 1 antibody
    • SGK antibody
    • SGK1 antibody
    • Sgk1 variant i3 antibody
    • SGK1_HUMAN antibody
    see all

Images

  • All lanes : Anti-SGK1 antibody (ab43606) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 4 : Human pancreas tissue lysate - total protein (ab29816)
    Lane 5 : Lung (Human) Tissue Lysate
    Lane 6 : Human brain tissue lysate - total protein (ab29466)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 49 kDa
    Observed band size: 49 kDa
    Additional bands at: 38 kDa, 51 kDa (possible isoform), 60 kDa (possible isoform), 95 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 5 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab43606 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • ICC/IF image of ab43606 stained HeLa cells. The cells were 100% methanol fixed (5 min) then permeabilised using 0.1% PBS-Triton and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to further permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab43606 at 5µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • Immunohistochemical analysis of human colon cancer tissue labeling SGK1 with ab43606 at 1/2000. A Goat Anti-rabbit HRP polymer was used as the secondary antibody. The tissue was fixed with Formaldehyde. Antigen retrieval was heat mediated.

    See Abreview

  • All lanes : Anti-SGK1 antibody (ab43606) at 1 µg/ml

    Lane 1 : Human BT549 (Breast carcinoma cell line) Whole Cell Lysate
    Lane 2 : T47D whole cell lysate (ab14899)
    Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 49 kDa
    Observed band size: 49 kDa


    Exposure time: 1 minute


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab43606 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

References

This product has been referenced in:

  • Wang Y  et al. Mechanism of Regulation of Big-Conductance Ca2+-Activated K+ Channels by mTOR Complex 2 in Podocytes. Front Physiol 10:167 (2019). Read more (PubMed: 30873046) »
  • Essig K  et al. Roquin targets mRNAs in a 3'-UTR-specific manner by different modes of regulation. Nat Commun 9:3810 (2018). Read more (PubMed: 30232334) »
See all 6 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rabbit Cell (heart)
Permeabilization
No
Specification
heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Apr 30 2018

Answer

Thank you for contacting us.

The good news first: both antibodies are suitable for your purpose. However, the ab59337 has actually been used with mouse stratium. Please see the attached reference.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Question
Answer

Thank you for contacting us.

The lab let me know that they haven't tested the antibodyto determine cross-reactivity with sgk2 and 3 but they blasted the immunogenagainst theHuman sequences and it only picked up sgk1.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Question
Answer

Thank you for contacting us.

Your credit note ID is **********

I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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Question
Answer

DISCOUNT CODE: ***
Expiration date: June 17th, 2012

I am very pleased to hear you would like to accept our offer and test ab43606 insheep. This code will give you 1 freeprimary antibodybefore the expiration date. To redeem this offer, please submit an Abreview forsheep and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More

Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab43606 with the order number 998916. To check the status of the order please contact our Customer Service team and reference this number. **Please note we discussed storing this antibody at 4 degrees C rather than -20 degrees C.** This free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (kidney)
Specification
kidney
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer pH6
Permeabilization
Yes - 1% TBST
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Dec 24 2010

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Intestinal adenoma)
Specification
Intestinal adenoma
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako Target Retrieval Solution
Permeabilization
No
Blocking step
5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C

Mr. Antibody Solutions

Verified customer

Submitted Jun 22 2009

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Colon Cancer)
Specification
Colon Cancer
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako Target Retrieval Solution
Permeabilization
No
Blocking step
5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C

Mr. Antibody Solutions

Verified customer

Submitted Jun 10 2009

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