NF-κB1 p105/p50 Antibody (YA6065)

Cat. No.: HY-P86373: Data Sheet COA
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NF-κB1 p105/p50 Antibody (YA6065) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to NF-κB1 p105/p50.

For research use only. We do not sell to patients.

Size	Stock	Quantity
10 μL	In-stock	Estimated Time of Arrival: December 31
50 μL	In-stock	Estimated Time of Arrival: December 31
100 μL	In-stock	Estimated Time of Arrival: December 31
250 μL	Get quote

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WB: Western Blot;
IHC-P: Immunohistochemistry-Paraffin;
IHC-F: Immunohistochemistry-Frozen;
ICC/IF: Immunocytochemistry/Immunofluorescence;
IF-Tissue: Immunofluorescence-Tissue;
mIHC: Multiplex Immunohistochemical;
IP: Immunoprecipitation;
ChIP: Chromatin Immunoprecipitation;
FC: Flow Cytometry;
ELISA: Enzyme Linked Immunosorbent Assay

Product Detail
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Background
Documentation
References

Description

NF-κB1 p105/p50 Antibody (YA6065) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to NF-κB1 p105/p50.

Host

Rabbit

Clonality

Monoclonal

Molecular Weight

Predicted band size: 50 kDa,105 kDa;

Observed band size: 50 kDa,120 kDa

Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.

Species Reactivity

Human, Mouse, Rat

SwissProt ID

P25799

Gene ID

4790 [NCBI]

Application &
Dilution Ratio

Application	Dilution Ratio
WB WB: Western Blot	1:2000-1:10000
IHC-P IHC-P: Immunohistochemistry-Paraffin	1:200-1:1000
ICC/IF ICC/IF: Immunocytochemistry/Immunofluorescence	1:200-1:1000
ELISA ELISA: Enzyme Linked Immunosorbent Assay	1:5000-1:20000
IP IP: Immunoprecipitation	1:50-1:200

Purity	Protein A	Conjugation	Non-conjugated
Modification	Unmodified	Isotype	IgG

Appearance

Liquid

Formulation

Supplied in PBS, 50% glycerol, 0.05% Proclin 300, 0.05%BSA

Storage & Stability

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping

Shipping with blue ice.

Verification Image

ALL WB IHC-P ICC

Western blot analysis of extracts from A431 (lane2(20μg), Hela (lane3(20μg), Jurkat (lane4(20μg) and Raji (lane5(20μg) using NF-κB1 p105/p50 Antibody (HY-P86373). Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/5000) and Loading control antibody (Beta Actin, HY-P80993, 1/10,000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HY-P8001 ,1/10,000) was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue using NF-κB1 p105/p50 Antibody (YA6065). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P86373, 1/150) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with neutral balsam.
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using NF-κB1 p105/p50 Antibody (YA6065). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P86373, 1/150) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with neutral balsam.
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue using NF-κB1 p105/p50 Antibody (YA6065). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P86373, 1/150) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with neutral balsam.
Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue using NF-κB1 p105/p50 Antibody (YA6065). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P86373, 1/150) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with neutral balsam.
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using NF-κB1 p105/p50 Antibody (YA6065). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P86373, 1/150) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with neutral balsam.
Immunohistochemical analysis of paraffin-embedded human lung cancer tissue using NF-κB1 p105/p50 Antibody (YA6065). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P86373, 1/150) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with neutral balsam.
Immunocytochemistry analysis of NIH-3T3 cells labeling NF-κB1 p105/p50 with NF-κB1 p105/p50 Antibody (HY-P86373) at 1:200 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with NF-κB1 p105/p50 Antibody (HY-P86373) at 1:200 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green） was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
Immunocytochemistry analysis of NIH-3T3 cells labeling NF-κB1 p105/p50 with NF-κB1 p105/p50 Antibody (HY-P86373) at 1:400 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with NF-κB1 p105/p50 Antibody (HY-P86373) at 1:400 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green） was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

Background

Function：NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes 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. Plays a role in the regulation of apoptosis. In a complex with MAP3K8, NFKB1/p105 represses MAP3K8-induced MAPK signaling; active MAP3K8 is released by proteasome-dependent degradation of NFKB1/p105; P105 is the precursor of the active p50 subunit (Nuclear factor NF-kappa-B p50 subunit) of the nuclear factor NF-kappa-B. Acts as a cytoplasmic retention of attached NF-kappa-B proteins by p105; Constitutes the active form, which associates with RELA/p65 to form the NF-kappa-B p65-p50 complex to form a transcription factor. Together with RELA/p65, 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; Isoform 3 (p98) (but not p84 or p105) acts as a transactivator of NF-kappa-B-regulated gene expression; Acts as an inhibitor of transactivation of p50 NF-kappa-B subunit, probably by sequestering it in the cytoplasm; Acts as an inhibitor of transactivation of p50 NF-kappa-B subunit, probably by sequestering it in the cytoplasm; Acts as an inhibitor of transactivation of p50 NF-kappa-B subunit, probably by sequestering it in the cytoplasm

Subcellular Localization：Cytoplasm; Nucleus; Cytoplasm; Cytoplasm; Nucleus; Cytoplasm; Nucleus

Expression：
Induction:By phorbol ester and TNF-alpha

Isoforms & Post-Translational Modification：P25799 has 7 isomers: P25799-1: 105615 Da (predicted); P25799-2: 84817 Da (predicted); P25799-3: 97413 Da (predicted); P25799-4: 39756 Da (predicted); P25799-5: 64782 Da (predicted); P25799-6: 56580 Da (predicted); P25799-7: 43983 Da (predicted).
Generation of the NF-kappa-B p50 (Nuclear factor NF-kappa-B p50 subunit) transcription factor takes place both cotranslationally and post-translationally via non-mutually exclusive mechanisms (By similarity). A cotranslational processing allows the production of both p50 and p105 (Nuclear factor NF-kappa-B p105 subunit) from a single NFKB1 mRNA (PubMed:9529257). While translation occurs, the particular unfolded structure after the GRR repeat region acts as a substrate for the proteasome, promoting degradation of the C-terminus (PubMed:9529257). The GRR acts as a proteasomal 'stop signal', protecting the region upstream of the GRR from degradation and promoting generation of p50 (PubMed:9529257). It is unclear if limited proteasome degradation during cotranslational processing depends on ubiquitination (PubMed:9529257). NF-kappa-B p50 is also generated post-translationally following ubiquitination by the KPC complex, leading to limited processing by the proteasome downstream of the GRR region, thereby generating p50 (By similarity);Phosphorylation at the C-terminus by IKBKB/IKKB acts as a signal for ubiquitination and promotes either complete degradation or processing to generate the NF-kappa-B p50 (Nuclear factor NF-kappa-B p50 subunit) (By similarity). Phosphorylation at Ser-910 primes p105 for proteolytic processing in response to TNF-alpha stimulation (By similarity). Phosphorylation at Ser-926, Ser-930 and Ser-935 are required for BTRC/BTRCP-mediated ubiquitination and proteolysis (By similarity). Phosphorylation at Ser-930 is also required for ubiquitination by the KPC complex and limited processing to generate NF-kappa-B p50 (Nuclear factor NF-kappa-B p50 subunit) (By similarity);Polyubiquitinated at multiple Lys residues in the C-terminus (By similarity). Polyubiquitinated by the SCF(FBXW11) and SCF(BTRC) complexes following phosphorylation at Ser-926, Ser-930 and Ser-935, leading to its complete degradation (By similarity). In contrast, polyubiquitination by the KPC complex following phosphorylation at Ser-930 leads to limited proteosomal processing and generation of the active NF-kappa-B p50 (Nuclear factor NF-kappa-B p50 subunit) (By similarity);S-nitrosylation of Cys-59 affects DNA binding;The covalent modification of cysteine by 15-deoxy-Delta12,14-prostaglandin-J2 is autocatalytic and reversible. It may occur as an alternative to other cysteine modifications, such as S-nitrosylation and S-palmitoylation

Subunit：Component of the NF-kappa-B p65-p50 complex (By similarity). Homodimer; component of the NF-kappa-B p50-p50 complex (By similarity). Component of the NF-kappa-B p105-p50 complex (By similarity). Component of the NF-kappa-B p50-c-Rel complex (By similarity). Component of a complex consisting of the NF-kappa-B p50-p50 homodimer and BCL3 (By similarity). Also interacts with MAP3K8 (By similarity). NF-kappa-B p50 subunit interacts with NCOA3 coactivator, which may coactivate NF-kappa-B dependent expression via its histone acetyltransferase activity (By similarity). Interacts with TSC22D3; this interaction prevents nuclear translocation and DNA-binding (By similarity). Interacts with SPAG9 and UNC5CL (By similarity). NFKB1/p105 interacts with CFLAR; the interaction inhibits p105 processing into p50 (By similarity). NFKB1/p105 forms a ternary complex with MAP3K8 and TNIP2 (By similarity). Interacts with GSK3B; the interaction prevents processing of p105 to p50 (By similarity). NFKB1/p50 interacts with NFKBIE (By similarity). NFKB1/p50 interacts with NFKBIZ (PubMed:11356851, PubMed:15241416). Nuclear factor NF-kappa-B p50 subunit interacts with NFKBID (PubMed:11931770). Directly interacts with MEN1 (By similarity). Interacts with HIF1AN (By similarity). Interacts with FEM1AA; interaction is direct (PubMed:18270204)

RRID

AB_3719138

Synonyms

NFKB1; Nuclear factor NF-kappa-B p105 subunit; DNA-binding factor KBF1; EBP-1; Nuclear factor of kappa light polypeptide gene enhancer in B-cells 1

Documentation

Select Batch:

Data Sheet (237 KB) SDS (252 KB)

COA (205 KB)

User Guide for Antibodies (1077 KB)

References

[1]. /biology-dictionary/pc-3-cell.html [Content Brief]