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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human RSK4 aa 1-100 (N terminal).
Database link: Q9UK32
This product is a recombinant rabbit monoclonal antibody.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
|WB||1/250 - 1/1000. Detects a band of approximately 90 kDa (predicted molecular weight: 84 kDa).|
|Flow Cyt||1/25 - 1/70.
ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|ICC/IF||1/100 - 1/1000.|
|IHC-P||1/50 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).
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 IRDye® 800CW Goat anti-Rabbit IgG (H + L) and IRDye® 680 Goat anti-Mouse IgG (H + L) secondary antibodies at 1:10,000 dilution for 1 hour at room temperature before imaging.
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.
Unpurified ab76117, at 1/50 dilution, staining RSK4 in paraffin-embedded human brain glioma tissue.
Immunofluorescent staining of SH-Sy5y cells using unpurified ab76117 at 1/100 dilution.