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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human IKB alpha (C terminal).
This product is a recombinant rabbit monoclonal antibody.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
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.
Our Abpromise guarantee covers the use of ab76429 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).|
ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
For unpurified use at 1/1000.
|IHC-P||1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).
For unpurified use at 1/100 - 1/250.
For unpurified use at 1/50 - 1/100.
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: IKB alpha knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lanes 1 - 3: Merged signal (red and green). Green - ab76429 observed at 39 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab76429 was shown to specifically react with IKB alpha in wild-type HAP1 cells as well as additional cross-reactive bands. No bands were observed when IKB alpha knockout cells were examined. Wild-type and IKB alpha knockout samples were subjected to SDS-PAGE. Ab76429 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/500 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
Immunohistochemical analysis of paraffin-embedded human colon tissue sections labelling IKB alpha with purified ab76429 at dilution of 1/500. Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
Immunocytochemistry/Immunofluorescence staining of HeLa cells labelling IKB alpha with purified ab76429 at a working dilution of 1/250. 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. DAPI was used as nuclear counterstain. The cells were fixed in 4% Paraformaldehyde and permeabilized using 0.1% Triton X-100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, rabbit primary antibody was used followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120). For negative control 2, ab7291 (mouse anti-tubulin) was used followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077).
Overlay histogram showing 4% paraformaldehyde fixed Jurkat (human acute T cell leukemia) cells labelling IKB alpha with purified ab76429 at dilution of 1/250. The secondary antibody used was Alexa Fluor® 488 goat-anti-rabbit IgG at dilution of 1/2000. A non-specific IgG antibody (rabbit monoclonal) was used as isotype control (black line). The blue line shows cells without incubation with primary antibody and secondary antibody.
Immunohistochemical analysis of Paraffin-embedded human colon tissue section labelling IKB alpha with ab76429 at 1/100 dilution.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"