Recombinant Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263)
![Western blot - Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263)](https://www.abcam.com/ps/products/305/ab305263/Images/ab305263-5-anti-nfkb-p105p50-antibody-1298ct792105117133-western-blot-wild-type-hap1human-chronic-myelogenous-leukemia-near-haploid-cell-line-whole-cell-lysate-20-ug.jpg "Western blot - Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [1298CT792.105.117.133] to NFkB p105 / p50
- Suitable for: WB, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133]
See all NFkB p105 / p50 primary antibodies -
Description
Mouse monoclonal [1298CT792.105.117.133] to NFkB p105 / p50 -
Host species
Mouse -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt (Intra)more details
Unsuitable for: ICC/IF or IP -
Species reactivity
Reacts with: Human
Does not react with: Mouse, Rat -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC:P: Human tonsil tissue. Human colon cancer. WB: Wild-type HAP1 whole cell lysate. HeLa, Daudi, Raji and THP-1 whole cell lysate. Human cerebellum, heart and kidney tissue lysate. Flow Cyt (Intra): HeLa and Raji cells.
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General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
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
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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
1298CT792.105.117.133 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab305263 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 |
1/1000. Predicted molecular weight: 105 kDa.
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| IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| Flow Cyt (Intra) |
1/100.
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| Notes |
|---|
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WB
1/1000. Predicted molecular weight: 105 kDa. |
|
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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Flow Cyt (Intra)
1/100. |
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
- Omim: 164011 Human
- SwissProt: P19838 Human
- Unigene: 618430 Human
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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 [1298CT792.105.117.133] (ab305263)All lanes : Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263) at 1/1000 dilution
Lane 1 : Wild-type HAP1(human chronic myelogenous leukemia near-haploid cell line), whole cell lysate
Lane 2 : NFkB p105 / p50 knockout HAP1, whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) at 1/10000 dilution
Predicted band size: 105 kDa
Observed band size: 50, 105 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.
False colour image of Western blot: Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263) staining at 1/1000 dilution, shown in green; Rabbit anti-GAPDH antibody [16891] (ab181602) loading control staining at 1/20000 dilution, shown in red.
In Western blot, ab305263 was shown to bind specifically to NFkB p105 / p50. A band was observed at 50/105 kDa in wild-type HAP1 cell lysates with no signal observed at this size in NFkB p105 / p50 knockout cell line. To generate this image, wild-type and NFkB p105 / p50 knockout HAP1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat Anti-Mouse IgG H&L (IRDye® 800CW) (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution. -
![Western blot - Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263)](https://www.abcam.com/ps/products/305/ab305263/Images/ab305263-6-anti-nfkb-p105p50-antibody-1298ct792105117133-western-blot-raji-human-burkitts-lymphoma-b-lymphocyte-whole-cell-lysate10-ug.jpg)
Western blot - Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263)All lanes : Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263) at 1/1000 dilution
Lane 1 : Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lane 2 : Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate
Lane 3 : THP-1 (human monocytic leukemia monocyte) whole cell lysate
Lane 4 : Human cerebellum tissue lysate
Lane 5 : Human heart tissue lysate
Lane 6 : Human kidney tissue lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Predicted band size: 105 kDa
Observed band size: 50, 105 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure times:
Lanes 1-3: 26 seconds
Lanes 4-6: 3 minutes -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263)](https://www.abcam.com/ps/products/305/ab305263/Images/ab305263-1-anti-nfkb-p105p50-antibody-1298ct792105117133-immunohistochemistry-human-tonsil.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263)Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling NFkB p105 / p50 with AB305263 at 1/2000 dilution (0.487 µg/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Positive staining on human tonsil.
The section was incubated with ab305263 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263)](https://www.abcam.com/ps/products/305/ab305263/Images/ab305263-2-anti-nfkb-p105p50-antibody-1298ct792105117133-immunohistochemistry-human-colon-cancer.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263)Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling NFkB p105 / p50 with AB305263 at 1/2000 dilution (0.487 µg/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Positive staining on human colon cancer.
The section was incubated with ab305263 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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![Flow Cytometry (Intracellular) - Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263)](https://www.abcam.com/ps/products/305/ab305263/Images/ab305263-3-anti-nfkb-p105p50-antibody-1298ct792105117133-flow-cytometry-hela.jpg)
Flow Cytometry (Intracellular) - Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263)Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling NFkB p105 / p50 with AB305263 at 1/100 dilution (1µg) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
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![Flow Cytometry (Intracellular) - Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263)](https://www.abcam.com/ps/products/305/ab305263/Images/ab305263-4-anti-nfkb-p105p50-antibody-1298ct792105117133-flow-cytometry-raji.jpg)
Flow Cytometry (Intracellular) - Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263)Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Raji (human Burkitt's lymphoma B lymphocyte) cells labeling NFkB p105 / p50 with AB305263 at 1/100 dilution (1µg) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
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![Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263)](https://www.abcam.com/ps/products/305/ab305263/Images/ab305263-496313-BenefitsofRecombinantantibodies.jpg)
Anti-NFkB p105 / p50 antibody [1298CT792.105.117.133] (ab305263)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab305263 has not yet been referenced specifically in any publications.