Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] (ab76302)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2294Y] to NF-kB p65 (phospho S536)
- Suitable for: Dot blot, WB, IP
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-NF-kB p65 (phospho S536) antibody [EP2294Y]
See all NF-kB p65 primary antibodies -
Description
Rabbit monoclonal [EP2294Y] to NF-kB p65 (phospho S536) -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IP HumanWB MouseRatHuman -
Immunogen
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human NF-kB p65 (phospho S536). The exact sequence is proprietary.
(Peptide available asab202905) -
Positive control
- WB: HeLa whole cell lysate treated with Calyculin A + TNF-alpha. C6 and NIH/3T3 treated with 100nM Calyculin A for 30 minutes whole cell lysate . IP: Daudi cell lysate treated with Calyculin A + TNF-alpha.
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General notes
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.
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
EP2294Y -
Isotype
IgG -
Research areas
Images
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ab76302 at 1/30 immunoprecipitating NF-kB p65 (phospho S536) in Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha.
Lane 1 (input): Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha (10µg)
Lane 2 (+): ab76302 + Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76302 in Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha.
For western blotting, ab76302 at 1/500 dilution (0.95 μg/ml) and VeriBlot for IP Detection Reagent (HRP)(ab131366) at 1/1000 dilution were used.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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All lanes : Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] (ab76302) at 1/1000 dilution
Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysate
Lane 2 : C6 (Rat glial tumor glial cell) treated with 100nM Calyculin A for 30 minutes whole cell lysate
Lane 3 : C6 (Rat glial tumor glial cell) treated with 100nM Calyculin A for 30 minutes, then the membrane treated with Alkaline Phosphatase for 1 hour
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: 65 kDa
Observed band size: 65 kDa
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All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with 100nM Calyculin A for 30 minutes whole cell lysate
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) treated with 100nM Calyculin A for 30 minutes, then the membrane treated with Alkaline Phosphatase for 1 hour
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: 65 kDa
Observed band size: 65 kDa
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All lanes : Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] (ab76302) at 1/1000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with Calyculin A and TNF-alpha 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: 65 kDa
Observed band size: 65 kDa
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Dot blot analysis of INF- kB p65 (phospho S536) phospho peptide (Lane 1) and NF- kB p65 non-phospho peptide (Lane 2) labeling NF-kB p65 (phospho S536) with ab76302 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG) (H+L) at 1/100 000 was used as the secondary antibody.
Blocking and diluting buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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