Recombinant Anti-SIK3 antibody [EPR23554-71] (ab255701)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23554-71] to SIK3
- Suitable for: ICC/IF, IP, WB, IHC-P, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-SIK3 antibody [EPR23554-71]
See all SIK3 primary antibodies -
Description
Rabbit monoclonal [EPR23554-71] to SIK3 -
Host species
Rabbit -
Specificity
This antibody does not react with Mouse and Rat species for IHC application.
This antibody does not react with Human species for ICC and FC application.
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Tested applications
Suitable for: ICC/IF, IP, WB, IHC-P, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Whole cell lysates of: U-87 MG (human glioblastoma-astrocytoma epithelial cell), Neuro-2a (mouse neuroblastoma neuroblast), C6 (rat glial tumor glial cell). Tissue lysates from: human brain, mouse brain, rat brain. IHC-P: human ovarian cancer, human bladder cancer. ICC/IF: Neuro-2a (mouse neuroblastoma neuroblast) cells. Flow cyt. (Intra.): Neuro-2a (mouse neuroblastoma neuroblast) cells.
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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.
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
EPR23554-71 -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab255701 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 |
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ICC/IF |
1/50.
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IP |
1/30.
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WB |
1/1000. Predicted molecular weight: 140 kDa.
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IHC-P |
1/100.
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Flow Cyt (Intra) |
1/50.
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Notes |
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ICC/IF
1/50. |
IP
1/30. |
WB
1/1000. Predicted molecular weight: 140 kDa. |
IHC-P
1/100. |
Flow Cyt (Intra)
1/50. |
Target
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Sequence similarities
Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. SNF1 subfamily.
Contains 1 protein kinase domain.
Contains 1 UBA domain. -
Post-translational
modificationsPhosphorylated at Thr-163 by STK11/LKB1 in complex with STE20-related adapter-alpha (STRADA) pseudo kinase and CAB39. -
Cellular localization
Cytoplasm. Locates to punctate structures within the cytoplasm on binding to YWHAZ. - Information by UniProt
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Database links
- Entrez Gene: 23387 Human
- Entrez Gene: 70661 Mouse
- Entrez Gene: 684112 Rat
- Omim: 614776 Human
- SwissProt: Q9Y2K2 Human
- SwissProt: Q6P4S6 Mouse
- Unigene: 167451 Human
- Unigene: 219459 Mouse
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Alternative names
- FLJ12240 antibody
- KIAA0999 antibody
- L19 antibody
see all
Images
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All lanes : Anti-SIK3 antibody [EPR23554-71] (ab255701) at 1/1000 dilution
Lane 1 : U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate
Lane 2 : NIH:OVCAR-3 (human ovary adenocarcinoma epithelial cell), whole cell lysate
Lane 3 : Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate
Lane 4 : C6 (rat glial tumor glial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 140 kDa
Observed band size: 73,69 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking / Dilution buffer and concentration: 5% NFDM/TBST
The expression profile/molecular weight observed is consistent with that described in the literature (PMID: 32542037).
Low expression: NIH:OVCAR-3 (PMID: PMID: 31762812) -
All lanes : Anti-SIK3 antibody [EPR23554-71] (ab255701) at 1/1000 dilution
Lane 1 : Human brain tissue lysate at 60 µg
Lane 2 : Mouse brain tissue lysate at 20 µg
Lane 3 : Rat brain tissue lysate at 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 140 kDa
Observed band size: 73,69 kDa why is the actual band size different from the predicted?Blocking buffer and concentration: 5% NFDM/TBST
Exposure time:
Lane 1: 3 minutes
Lane 2 & 3: 26 secondsThe expression profile/molecular weight observed is consistent with that described in the literature (PMID: 32542037).
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Immunohistochemical analysis of paraffin-embedded Human ovarian cancer tissue labelling SIK3 with ab255701 at 1/100 (5.85 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in human ovarian cancer (PMID: 21399663). The section was incubated with ab255701 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
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Immunohistochemical analysis of paraffin-embedded Human bladder cancer tissue labelling SIK3 with ab255701 at 1/100 (5.85 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in human bladder cancer. The section was incubated with ab255701 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
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells lebelling SIK3 with ab255701 at 1/50 (11.7 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/mL) dilution (Green). Confocal image showing cytoplasmic staining in Neuro-2a cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2ug/mL) dilution.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (Mouse neuroblastoma neuroblast) cells labelling SIK3 with ab255701 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
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SIK3 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate 10 ug with ab255701 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab255701 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Mouse brain tissue lysate 10 ug
Lane 2: ab255701 IP in Mouse brain tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab255701 in Mouse brain tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 41 seconds
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab255701 has not yet been referenced specifically in any publications.