Recombinant Anti-NFkB p105 / p50 antibody [E381] (ab32360)
![Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)](https://www.abcam.com/ps/products/32/ab32360/Images/ab32360-268407-anti-nfkb-p105-p50-antibody-e381-western-blot.jpg "Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E381] to NFkB p105 / p50
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-NFkB p105 / p50 antibody [E381]
See all NFkB p105 / p50 primary antibodies -
Description
Rabbit monoclonal [E381] to NFkB p105 / p50 -
Host species
Rabbit -
Specificity
This antibody will detect both forms: p50 and p105. -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, MCF-7 and PC-12 cell lysates. IHC-P: human prostate carcinoma and bladder carcinoma tissues.
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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.
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.
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, 0.05% BSA -
Concentration information loading... -
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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Assay kits
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32360 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 |
|---|---|---|
| WB | (11) |
1/1000 - 1/10000. Detects a band of approximately 50, 105 kDa (predicted molecular weight: 50 kDa).
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| IHC-P | (5) |
1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| Notes |
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WB
1/1000 - 1/10000. Detects a band of approximately 50, 105 kDa (predicted molecular weight: 50 kDa). |
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IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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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Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)All lanes : Anti-NFkB p105 / p50 antibody [E381] (ab32360)
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : NFkB p105 / p50 knockout HAP1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 50 kDaLanes 1 - 4: Merged signal (red and green). Green - ab32360 observed at 120, 50 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32360 was shown to specifically react with NFkB p105 / p50 when NFkB p105 / p50 knockout samples were used. Wild-type and NFkB p105 / p50 knockout samples were subjected to SDS-PAGE. ab32360 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB p105 / p50 antibody [E381] (ab32360)](https://www.abcam.com/ps/products/32/ab32360/Images/ab32360-250267-anti-nfkb-p105-p50-antibody-e381-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB p105 / p50 antibody [E381] (ab32360)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue labelling NFkB p105 / p50 with purified ab32360 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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![Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)](https://www.abcam.com/ps/products/32/ab32360/Images/ab32360-409568-anti-nfkb-p105-p50-antibody-e381-western-blot-hela-edited-lysates.jpg)
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)All lanes : Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : NFKB1 CRISPR/Cas9 edited HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab32360 observed at 105 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32360 was shown to react with NFkB p105 / p50 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab264823 (CRISPR/Cas9 edited cell lysate ab257003) lane below 105kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and NFKB1 CRISPR/Cas9 HeLa HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32360 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)](https://www.abcam.com/ps/products/32/ab32360/Images/ab32360-250255-anti-nfkb-p105-p50-antibody-e381-western-blot.jpg)
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/10000 dilution (purified) + HeLa cell lysate at 20 µg
Secondary
Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 50 kDa
Observed band size: 105,50 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)](https://www.abcam.com/ps/products/32/ab32360/Images/ab32360-250256-anti-nfkb-p105-p50-antibody-e381-western-blot.jpg)
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/50000 dilution (purified) + MCF-7 cell lysate at 10 µg
Secondary
Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 50 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?
Additional bands at: 50 kDa (possible non-specific binding)Blocking and dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)](https://www.abcam.com/ps/products/32/ab32360/Images/ab32360-250257-anti-nfkb-p105-p50-antibody-e381-western-blot.jpg)
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)All lanes : Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/10000 dilution (purified)
Lane 1 : PC-12 cell lysate
Lane 2 : NIH/3T3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 50 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?
Additional bands at: 50 kDa (possible non-specific binding)Blocking and dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)](https://www.abcam.com/ps/products/32/ab32360/Images/ab32360-44553-anti-nfkb-p105-p50-antibody-e381-western-blot.jpg)
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/5000 dilution (unpurified) + HeLa cell lysate
Predicted band size: 50 kDa
Observed band size: 105,50 kDa why is the actual band size different from the predicted? -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB p105 / p50 antibody [E381] (ab32360)](https://www.abcam.com/ps/products/32/ab32360/Images/ab32360-44552-anti-nfkb-p105-p50-antibody-e381-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB p105 / p50 antibody [E381] (ab32360)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling NFkB p105 / p50 with unpurified ab32360 at a dilution of 1/250.
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![Anti-NFkB p105 / p50 antibody [E381] (ab32360)](https://www.abcam.com/ps/products/32/ab32360/Images/ab32360-12-benefits-of-recombinant-antibodies.png)
Anti-NFkB p105 / p50 antibody [E381] (ab32360)
Datasheets and documents
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SDS download
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Datasheet download
References (161)
ab32360 has been referenced in 161 publications.
- Cao W et al. Naringin Targets NFKB1 to Alleviate Oxygen-Glucose Deprivation/Reoxygenation-Induced Injury in PC12 Cells Via Modulating HIF-1a/AKT/mTOR-Signaling Pathway. J Mol Neurosci 71:101-111 (2021). PubMed: 32557145
- Zou XS et al. Pomelo peel oil alleviates cerebral NLRP3 inflammasome activation in a cardiopulmonary resuscitation rat model. Exp Ther Med 21:233 (2021). PubMed: 33603841
- Huang D et al. Casticin protected against neuronal injury and inhibited the TLR4/NF-?B pathway after middle cerebral artery occlusion in rats. Pharmacol Res Perspect 9:e00752 (2021). PubMed: 33704926
- Zhang L et al. The anti-breast cancer property of physcion via oxidative stress-mediated mitochondrial apoptosis and immune response. Pharm Biol 59:303-310 (2021). PubMed: 33715588
- Zhou Z et al. LncRNA XIST promotes inflammation by downregulating GRa expression in the adenoids of children with OSAHS. Exp Ther Med 21:500 (2021). PubMed: 33791009