Recombinant Anti-RSK4 antibody [EP1982Y] (ab76117)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1982Y] to RSK4
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-RSK4 antibody [EP1982Y]
See all RSK4 primary antibodies -
Description
Rabbit monoclonal [EP1982Y] to RSK4 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IF, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human RSK4 aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: Q9UK32 -
Positive control
- SH-SY5Y cell lysate. Human brain glioma tissue. SH-Sy5y 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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
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
EP1982Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab76117 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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Flow Cyt (Intra) |
1/25 - 1/70.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/250 - 1/1000. Detects a band of approximately 90 kDa (predicted molecular weight: 84 kDa).
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ICC/IF |
1/100 - 1/1000.
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IHC-P |
1/50 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/25 - 1/70. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/250 - 1/1000. Detects a band of approximately 90 kDa (predicted molecular weight: 84 kDa). |
ICC/IF
1/100 - 1/1000. |
IHC-P
1/50 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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Function
Serine/threonine kinase that may play a role in mediating the growth-factor and stress induced activation of the transcription factor CREB. -
Sequence similarities
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. S6 kinase subfamily.
Contains 1 AGC-kinase C-terminal domain.
Contains 2 protein kinase domains. -
Post-translational
modificationsAutophosphorylated on Ser-389, as part of the activation process. - Information by UniProt
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Database links
- Entrez Gene: 27330 Human
- Entrez Gene: 67071 Mouse
- Entrez Gene: 317203 Rat
- Omim: 300303 Human
- SwissProt: Q9UK32 Human
- SwissProt: Q7TPS0 Mouse
- Unigene: 234478 Human
- Unigene: 37617 Mouse
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Alternative names
- 90 kDa ribosomal protein S6 kinase 6 antibody
- KS6A6_HUMAN antibody
- p90 RSK 6 antibody
see all
Images
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Lane 1: Wild-type HAP1 cell lysate (40 µg)
Lane 2: RSK4 knockout HAP1 cell lysate (40 µg)
Lane 3: MCF7 cell lysate (20 µg)
Lane 4: HEK293 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab76117 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.ab76117 was shown to recognize RSK4 when RSK4 knockout samples were used, along with additional cross-reactive bands. Wild-type and RSK4 knockout samples were subjected to SDS-PAGE. Ab76117 and ab8245 (loading control to GAPDH) were diluted at 1/250 and 1/10,000 dilution respectively and incubated overnight at 4C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-RSK4 antibody [EP1982Y] (ab76117) at 1/1000 dilution (purified)
Lane 1 : SH-SY5Y cell lysate
Lane 2 : Caco-2 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 84 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Immunofluorescence staining of HEK293 cells with purified ab76117 at a working dilution of 1/1000, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab76117 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
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Immunohistochemical staining of paraffin embedded rat kidney with purified ab76117 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Immunohistochemical staining of paraffin embedded mouse cerebral cortex with purified ab76117 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Immunohistochemical staining of paraffin embedded human kidney with purified ab76117 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Overlay histogram showing Neuro-2a cells fixed in 2% PFA and stained with purified ab76117 at a dilution of 1 in 70 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
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Anti-RSK4 antibody [EP1982Y] (ab76117) at 1/500 dilution (unpurified) + SH-SY5Y cell lysate at 10 µg
Secondary
HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 84 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted? -
Unpurified ab76117, at 1/50 dilution, staining RSK4 in paraffin-embedded human brain glioma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunofluorescent staining of SH-Sy5y cells using unpurified ab76117 at 1/100 dilution.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (7)
ab76117 has been referenced in 7 publications.
- Hu C et al. Raltitrexed regulates proliferation and apoptosis of HGC-27 cells by upregulating RSK4. BMC Pharmacol Toxicol 23:65 (2022). PubMed: 36031631
- Mei Y et al. Overexpression of RSK4 reverses doxorubicin resistance in human breast cancer cells via PI3K/AKT signalling pathway. J Biochem 167:603-611 (2020). PubMed: 31960922
- Liang H et al. MiR-548d-3p Promotes Gastric Cancer by Targeting RSK4. Cancer Manag Res 12:13325-13337 (2020). PubMed: 33380838
- Hu C et al. Effect of RSK4 on Biological Characteristics of Gastric Cancer. Cancer Manag Res 12:611-619 (2020). PubMed: 32095082
- Huo H et al. RSK4 inhibits breast cancer cell proliferation and invasion in vitro, and is correlated with estrogen receptor upregulation in breast cancer. Oncol Rep 42:2777-2787 (2019). PubMed: 31545499
- Li Q et al. Estradiol promotes the progression of ER+ breast cancer through methylation-mediated RSK4 inactivation. Onco Targets Ther 12:5907-5916 (2019). PubMed: 31413588
- Sun Y et al. Basic anatomy and tumor biology of the RPS6KA6 gene that encodes the p90 ribosomal S6 kinase-4. Oncogene : (2012). WB ; Human . PubMed: 22614021