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AB225633

Anti-SIK3 (phospho T469) antibody [EPR20877-104]

  • BOND RX™ Validated
  • 20ul selling size
  • RabMAb
  • Recombinant
  • What is this?

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(1 Publication)

Rabbit Recombinant Monoclonal SIK3 phospho T469 antibody. Suitable for ICC/IF, Flow Cyt (Intra), Dot, WB, IHC-P and reacts with Mouse, Transfected cell line - Human samples. Cited in 1 publication.

View Alternative Names

Kiaa0999, Qsk, Sik3, Serine/threonine-protein kinase SIK3, Salt-inducible kinase 3, Serine/threonine-protein kinase QSK, SIK-3

6 Images
Flow Cytometry (Intracellular) - Anti-SIK3 (phospho T469) antibody [EPR20877-104] (AB225633)
  • Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SIK3 (phospho T469) antibody [EPR20877-104] (AB225633)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (Human embryonic kidney epithelial cell) cells transfected with flag-tagged SIK3 WT expression vector cells labelling SIK3 (phospho T469) with ab225633 at 1/500 dilution (0.1ug) (Right) compared with a Rabbit monoclonal IgG isotype control (ab172730) (Left). A Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

Immunocytochemistry/ Immunofluorescence - Anti-SIK3 (phospho T469) antibody [EPR20877-104] (AB225633)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SIK3 (phospho T469) antibody [EPR20877-104] (AB225633)

Immunocytochemical analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells labelling SIK3 (phospho T469) with ab225633 at 1/100 (5.22 ug/mL) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody (ab150077) at 1/1000 (2 ug/mL) dilution (Green). Confocal image showing cytoplasmic staining in 293T cells transfected with flag-tagged SIK3 WT expression vector, and no staining in 293T cells transfected with flag-tagged SIK3 T469A expression vector. Anti-FLAG mouse monoclonal antibody was used to counterstain tubulin at 1/500 (2 ug/mL) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 (2 ug/mL) dilution.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIK3 (phospho T469) antibody [EPR20877-104] (AB225633)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIK3 (phospho T469) antibody [EPR20877-104] (AB225633)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling SIK3 (phospho T469) with ab225633 at 1/1000 (0.522 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on mouse cerebral cortex without alkaline phosphatase treatment (image A). No signal was detected when tissues were treated with alkaline phosphatase (image B). The section was incubated with ab225633 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Western blot - Anti-SIK3 (phospho T469) antibody [EPR20877-104] (AB225633)
  • WB

Lab

Western blot - Anti-SIK3 (phospho T469) antibody [EPR20877-104] (AB225633)

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

Data was kindly provided by Dr. Qinghua Liu, NIBS.

The 1st Ab working concentration was 5 ug/ml.

All lanes:

Western blot - Anti-SIK3 (phospho T469) antibody [EPR20877-104] (ab225633) at 0.522 mg/mL

All lanes:

Mouse cortex tissue lysate at 4 µg

Predicted band size: 140 kDa

Observed band size: 150 kDa,70 kDa

false

Western blot - Anti-SIK3 (phospho T469) antibody [EPR20877-104] (AB225633)
  • WB

Lab

Western blot - Anti-SIK3 (phospho T469) antibody [EPR20877-104] (AB225633)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTThis blot was developed using a higher sensitivity ECL substrate.

Exposure time : 114 seconds

All lanes:

Western blot - Anti-SIK3 (phospho T469) antibody [EPR20877-104] (ab225633) at 1/5000 dilution

Lane 1:

Mouse hippocampus tissue lysate (Untreated membrane) at 10 µg

Lane 2:

Mouse hippocampus tissue lysate (Phosphatase treated membrane) at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 140 kDa

Observed band size: 70 kDa

false

Dot Blot - Anti-SIK3 (phospho T469) antibody [EPR20877-104] (AB225633)
  • Dot

Supplier Data

Dot Blot - Anti-SIK3 (phospho T469) antibody [EPR20877-104] (AB225633)

Dot blot analysis of SIK3 (phospho T469) using ab225633 at 1/1000 (0.522 ug/mL) dilution followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution.

Lane 1 : SIK3 (phospho T469) peptide

Lane 2 : SIK3 non-phospho peptide

Exposure time : 3 minutes.

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

  • Carrier free

    Anti-SIK3 (phospho T469) antibody [EPR20877-104] - BSA and Azide free

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR20877-104

Isotype

IgG

Carrier free

No

Reacts with

Mouse

Applications

Dot, WB, ICC/IF, IHC-P, Flow Cyt (Intra)

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "IHCFr" : {"fullname" : "Immunohistochemistry (Frozen sections)", "shortname":"IHC-Fr"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "1/5000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "1/1000", "IHCP-species-notes": "<p></p>", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" }, "Mouse": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/100", "ICCIF-species-notes": "<p></p>", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "FlowCytIntra-species-checked": "guaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "1/1000", "Dot-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/5000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/1000", "IHCP-species-notes": "<p></p>", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "<p></p>" }, "Rat": { "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "1/5000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "1/1000", "IHCP-species-notes": "<p></p>", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" }, "Transfected cell line - Human": { "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/460", "FlowCytIntra-species-notes": "<p></p>", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" } } }
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Product details

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

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

SIK3 also known as salt-inducible kinase 3 is a serine/threonine-protein kinase with a molecular mass of approximately 140 kDa. This protein is expressed in many tissues including liver muscle and pancreas. Mechanically SIK3 phosphorylates key substrates involved in the regulation of various cellular functions enhancing or inhibiting their activity. Its action often involves regulating metabolism cytokine signaling and cellular growth processes.
Biological function summary

SIK3 participates in the regulation of glucose metabolism and energy homeostasis. It acts as part of larger complexes that control transcription factors and enzymes related to metabolic processes. These complexes help respond to cellular signals and external stress adjusting cellular functions to maintain balance. It influences the activity of enzymes and transcriptional regulators involved in lipid and carbohydrate metabolism playing a role in energy balance maintenance.

Pathways

SIK3 influences gluconeogenesis and insulin signaling pathways. It is closely connected with proteins such as AMPK (AMP-activated protein kinase) regulating energy sensor mechanisms and metabolic stress responses. These pathways are critical in controlling glucose production and utilization which affect overall energy homeostasis in the body.

SIK3's dysfunction links to metabolic disorders like type 2 diabetes and obesity. Altered SIK3 activity affects insulin signaling which can contribute to impaired glucose regulation. It also connects to the protein LKB1 (liver kinase B1) in metabolic disease contexts as both play roles in regulating energy metabolism and maintaining glucose levels. Understanding SIK3's role provides insights for targeting metabolic diseases through therapeutic strategies.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Positive regulator of mTOR signaling that functions by triggering the degradation of DEPTOR, an mTOR inhibitor (By similarity). Required for chondrocyte hypertrophy during skeletogenesis (PubMed : 22318228). Negatively regulates cAMP signaling pathway possibly by acting on CRTC2/TORC2 and CRTC3/TORC3 (By similarity). Prevents HDAC4 translocation to the nucleus (PubMed : 22318228).
See full target information Sik3 phospho T469
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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

International journal of biological sciences 20:312-330 PubMed38164184

2024

Targeting phosphorylation circuits on CREB and CRTCs as the strategy to prevent acquired skin hyperpigmentation.

Applications

Unspecified application

Species

Unspecified reactive species

Song-Hee Kim,Changseon Na,Cheng-Yong Yun,Jun Gu Kim,Seung Tae Baek,Hyun Jin An,Jae Duk Lee,Seung Wha Lee,Jae-Kyung Jung,Bang Yeon Hwang,Sang-Bae Han,Youngoo Kim
View all publications
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Product promise

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