Rabbit Recombinant Monoclonal IKB alpha antibody. Suitable for IHC-P, WB, ICC/IF and reacts with Human, Rat, Mouse samples. Cited in 32 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IHC-P | Flow Cyt | WB | ICC/IF | |
---|---|---|---|---|
Human | Tested | Not recommended | Tested | Tested |
Mouse | Expected | Not recommended | Tested | Expected |
Rat | Expected | Not recommended | Tested | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/500 | Notes For unpurified use at 1/100 - 1/250. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/1000. |
Species Rat | Dilution info - | Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/1000. |
Species Human | Dilution info - | Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/1000. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1/500 - 1/10000 | Notes - |
Species Mouse | Dilution info 1/500 - 1/10000 | Notes - |
Species Human | Dilution info 1/500 - 1/10000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/250 | Notes For unpurified use at 1/50 - 1/100. |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
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Inhibits the activity of dimeric NF-kappa-B/REL complexes by trapping REL (RELA/p65 and NFKB1/p50) dimers in the cytoplasm by masking their nuclear localization signals (PubMed:1493333, PubMed:36651806, PubMed:7479976). On cellular stimulation by immune and pro-inflammatory responses, becomes phosphorylated promoting ubiquitination and degradation, enabling the dimeric RELA to translocate to the nucleus and activate transcription (PubMed:7479976, PubMed:7628694, PubMed:7796813, PubMed:7878466).
IKBA, MAD3, NFKBI, MAD3, NFKBI, IKBA, NFKBIA, NF-kappa-B inhibitor alpha, I-kappa-B-alpha, Major histocompatibility complex enhancer-binding protein MAD3, IkB-alpha, IkappaBalpha
Rabbit Recombinant Monoclonal IKB alpha antibody. Suitable for IHC-P, WB, ICC/IF and reacts with Human, Rat, Mouse samples. Cited in 32 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EP697
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), used at a dilution of 1/1000. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120). For negative control 2, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (mouse anti-tubulin) was used followed by an Alexa Fluor® 488 goat anti-rabbit antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077).
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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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.
All lanes: Western blot - Anti-IKB alpha antibody [EP697] (ab76429)
Predicted band size: 35 kDa
Blocking/Diluting buffer 5% NFDM /TBST
All lanes: Western blot - Anti-IKB alpha antibody [EP697] (ab76429) at 1/1000 dilution
Lane 1: Jurkat (human acute T cell leukemia) whole cell lysate at 15 µg
Lane 2: HeLa (human cervix adenocarcinoma) whole cell lysate at 15 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 35 kDa
Observed band size: 39 kDa
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.
Blocking/Diluting buffer 5% NFDM /TBST
All lanes: Western blot - Anti-IKB alpha antibody [EP697] (ab76429) at 1/10000 dilution
Lane 1: RAW264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate at 15 µg
Lane 2: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 15 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 35 kDa
Observed band size: 39 kDa
Immunohistochemical analysis of Paraffin-embedded human colon tissue section labelling IKB alpha with ab76429 at 1/100 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
All lanes: Western blot - Anti-IKB alpha antibody [EP697] (ab76429) at 1/1000 dilution
Lane 1: HeLa cell lysates at 10 µg
Lane 2: Jurkat cell lysates at 10 µg
All lanes: goat anti-rabbit HRP at 1/1000 dilution
Predicted band size: 35 kDa
Observed band size: 36 kDa
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