Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)
![Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)](https://www.abcam.com/ps/products/183/ab183559/Images/ab183559-456775-anti-nf-kb-p65-phospho-s276-antibody-epr17622-western-blot-hela-human.jpg "Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17622] to NF-kB p65 (phospho S276)
- Suitable for: ICC/IF, IP, WB
- Reacts with: Human
Overview
-
Product name
Anti-NF-kB p65 (phospho S276) antibody [EPR17622]
See all NF-kB p65 primary antibodies -
Description
Rabbit monoclonal [EPR17622] to NF-kB p65 (phospho S276) -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIP HumanWB Human
-
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: Lysates of HeLa cells treated with Calyculin A (100ng/ml, 30min), then TNF-a (20ng/ml, 5min). ICC/IF: HeLa cells treated with Calyculin A (100ng/ml, 10min), then TNF-a (20ng/ml, 5min). IP: HeLa whole cell lysate treated with Calyculin A (100ng/ml, 10min), then TNF-a (20ng/ml, 5min).
-
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
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
EPR17622 -
Isotype
IgG -
Research areas
Images
-
Western blot - Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)All lanes : Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559) at 1/1000 dilution
Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa whole cell lysate treated with 100ng/ml Calyculin A ab141784 for 30 minutes, then treated with 20ng/ml TNF-a for 5 minutes
Lane 3 : HeLa whole cell lysate treated with 100ng/ml Calyculin A ab141784 for 30 minutes, then treated with 20ng/ml TNF-a for 5 minutes, then treated with Alkaline Phosphatase for 1 hour
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 60 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesNegative control: Anti-NF-kB p65 antibody, ab32536
Blocking/Dilution buffer: 5% NFDM/TBST.
Phosphorylation of NF-kB p65 at S276 can be induced by TNF-a treatment according to the literature (PMID: 21795584).
-
![Immunocytochemistry/ Immunofluorescence - Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)](https://www.abcam.com/ps/products/183/ab183559/Images/ab183559-269448-anti-nf-kb-p65-phospho-s276-antibody-epr17622-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling NF-kB p65 (phospho S276) with ab183559 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
The expression increased on HeLa cells after treatment with Calyculin A (ab141784 100ng/ml, 10min) then TNF-a (20ng/ml, 5min).
The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
-
![Immunoprecipitation - Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)](https://www.abcam.com/ps/products/183/ab183559/Images/ab183559-269445-anti-nf-kb-p65-phospho-s276-antibody-epr17622-immunoprecipitation.jpg)
Immunoprecipitation - Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)NF-kB p65 (phospho S276) was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates, treated with 100ng/ml Calyculin A (ab141784) for 10min, then 20ng/ml TNA-a for 5min, with ab183559 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab183559 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HeLa whole cell lysate treated with 100ng/ml CA for 10min, then 20ng/ml TNA-a for 5min, 10 μg (Input).
Lane 2: ab183559 IP in HeLa whole cell lysate treated with 100ng/ml CA for 10min, then 20ng/ml TNA-a for 5min.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab183559 in HeLa whole cell lysate treated with 100ng/ml CA for 10min, then 20ng/ml TNA-a for 5min.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
-
![Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)](https://www.abcam.com/ps/products/183/ab183559/Images/ab183559-6-benefits-of-recombinant-antibodies.png)
Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)
