Recombinant HRP Anti-NFkB p105 / p50 antibody [E381] (ab195854)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- HRP Rabbit monoclonal [E381] to NFkB p105 / p50
- Suitable for: WB
- Knockout validated
- Reacts with: Human
- Conjugation: HRP
Related conjugates and formulations
Overview
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Product name
HRP Anti-NFkB p105 / p50 antibody [E381]
See all NFkB p105 / p50 primary antibodies -
Description
HRP Rabbit monoclonal [E381] to NFkB p105 / p50 -
Host species
Rabbit -
Conjugation
HRP -
Tested applications
Suitable for: WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa whole cell lysate.
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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.
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. Store In the Dark. -
Storage buffer
pH: 7.40
Preservative: 0.1% Proclin 300 Solution
Constituents: 30% Glycerol (glycerin, glycerine), 1% BSA, PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E381 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab195854 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/2000. Detects a band of approximately 50, 105 kDa (predicted molecular weight: 50 kDa).
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Notes |
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WB
1/2000. Detects a band of approximately 50, 105 kDa (predicted molecular weight: 50 kDa). |
Target
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Function
NF-kappa-B is a pleiotropic transcription factor which is present in almost all cell types and is involved in many biological processed such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. NF-kappa-B heterodimeric p65-p50 and RelB-p50 complexes are transcriptional activators. The NF-kappa-B p50-p50 homodimer is a transcriptional repressor, but can act as a transcriptional activator when associated with BCL3. NFKB1 appears to have dual functions such as cytoplasmic retention of attached NF-kappa-B proteins by p105 and generation of p50 by a cotranslational processing. The proteasome-mediated process ensures the production of both p50 and p105 and preserves their independent function, although processing of NFKB1/p105 also appears to occur post-translationally. p50 binds to the kappa-B consensus sequence 5'-GGRNNYYCC-3', located in the enhancer region of genes involved in immune response and acute phase reactions. In a complex with MAP3K8, NFKB1/p105 represses MAP3K8-induced MAPK signaling; active MAP3K8 is released by proteasome-dependent degradation of NFKB1/p105. -
Sequence similarities
Contains 7 ANK repeats.
Contains 1 death domain.
Contains 1 RHD (Rel-like) domain. -
Domain
The C-terminus of p105 might be involved in cytoplasmic retention, inhibition of DNA-binding, and transcription activation.
Glycine-rich region (GRR) appears to be a critical element in the generation of p50. -
Post-translational
modificationsWhile translation occurs, the particular unfolded structure after the GRR repeat promotes the generation of p50 making it an acceptable substrate for the proteasome. This process is known as cotranslational processing. The processed form is active and the unprocessed form acts as an inhibitor (I kappa B-like), being able to form cytosolic complexes with NF-kappa B, trapping it in the cytoplasm. Complete folding of the region downstream of the GRR repeat precludes processing.
Phosphorylation at 'Ser-903' and 'Ser-907' primes p105 for proteolytic processing in response to TNF-alpha stimulation. Phosphorylation at 'Ser-927' and 'Ser-932' are required for BTRC/BTRCP-mediated proteolysis.
Polyubiquitination seems to allow p105 processing.
S-nitrosylation of Cys-61 affects DNA binding. -
Cellular localization
Nucleus. Cytoplasm. Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor. - Information by UniProt
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Database links
- Entrez Gene: 4790 Human
- Entrez Gene: 18033 Mouse
- Entrez Gene: 81736 Rat
- Omim: 164011 Human
- SwissProt: P19838 Human
- SwissProt: P25799 Mouse
- SwissProt: Q63369 Rat
- Unigene: 618430 Human
see all -
Alternative names
- DKFZp686C01211 antibody
- DNA binding factor KBF1 antibody
- DNA binding factor KBF1 EBP1 antibody
see all
Images
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All lanes : HRP Anti-NFkB p105 / p50 antibody [E381] (ab195854) at 1/2000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Nfkb1 (NFkB p105 / p50) knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 50 kDa
Observed band size: 105,50 kDa why is the actual band size different from the predicted?
Exposure time: 20 minutesab195854 was shown to recognize NFkB p105 / p50 in wild-type HAP1 cells as signal was lost at the expected MW in Nfkb1 (NFkB p105 / p50) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Nfkb1 (NFkB p105 / p50) knockout samples were subjected to SDS-PAGE. Ab195854 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.
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HRP Anti-NFkB p105 / p50 antibody [E381] (ab195854) at 1/2000 dilution +
HeLa whole cell lysate (ab150035) at 10 µg
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 105,50 kDa why is the actual band size different from the predicted?
Exposure time: 20 minutesThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab195854 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (3)
ab195854 has been referenced in 3 publications.
- Chen Z et al. Two-layer regulation of TRAF6 mediated by both TLR4/NF-kB signaling and miR-589-5p increases proinflammatory cytokines in the pathology of severe acute pancreatitis. Am J Transl Res 12:2379-2395 (2020). PubMed: 32655778
- Cartwright TN et al. HDAC1 interacts with the p50 NF-?B subunit via its nuclear localization sequence to constrain inflammatory gene expression. Biochim Biophys Acta Gene Regul Mech 1861:962-970 (2018). PubMed: 30496041
- Qian Y et al. Lactobacillus casei Strain Shirota Enhances the In Vitro Antiproliferative Effect of Geniposide in Human Oral Squamous Carcinoma HSC-3 Cells. Molecules 23:N/A (2018). PubMed: 29751513