Antibodies, Proteins, Kits and Reagents for Life Science

304 886 ₸ Product size

100 µl 304 886 ₸

Order now and get it on Tuesday March 02, 2021

Anti-IKB alpha antibody [EP697] (ab76429)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP697] to IKB alpha
Suitable for: WB, IHC-P, ICC/IF
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-IKB alpha antibody [EP697]
See all IKB alpha primary antibodies
Description

Rabbit monoclonal [EP697] to IKB alpha
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Human
IHC-P	Human
WB	Mouse Human

See all applications and species data

Immunogen

Synthetic peptide within Human IKB alpha (C terminal). The exact sequence is proprietary.
Positive control
- HeLa, PC-12, RAW264.7 and Jurkat cell lysates; human kidney and colon tissue.
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
For more information see here.
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EP697
Isotype

IgG
Research areas

Western blot - Anti-IKB alpha antibody [EP697] (ab76429)

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.
Western blot - Anti-IKB alpha antibody [EP697] (ab76429)

All lanes : Anti-IKB alpha antibody [EP697] (ab76429) at 1/1000 dilution

Lane 1 : Jurkat (human acute T cell leukemia) whole cell lysate
Lane 2 : HeLa (human cervix adenocarcinoma) whole cell lysate

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 36 kDa
Observed band size: 39 kDa
why is the actual band size different from the predicted?

Blocking/Diluting buffer 5% NFDM /TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKB alpha antibody [EP697] (ab76429)

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 - Anti-IKB alpha antibody [EP697] (ab76429)

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).
Western blot - Anti-IKB alpha antibody [EP697] (ab76429)

All lanes : Anti-IKB alpha antibody [EP697] (ab76429) at 1/10000 dilution

Lane 1 : RAW264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate
Lane 2 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 36 kDa
Observed band size: 39 kDa why is the actual band size different from the predicted?

Blocking/Diluting buffer 5% NFDM /TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKB alpha antibody [EP697] (ab76429)

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.
Western blot - Anti-IKB alpha antibody [EP697] (ab76429)

All lanes : Anti-IKB alpha antibody [EP697] (ab76429) at 1/1000 dilution

Lane 1 : HeLa cell lysates
Lane 2 : Jurkat cell lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : goat anti-rabbit HRP at 1/1000 dilution

Predicted band size: 36 kDa
Observed band size: 36 kDa
Anti-IKB alpha antibody [EP697] (ab76429)

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