| Applications: |
CLIA |
| Reactivity: |
Rat |
| Note: |
FOR SCIENTIFIC EDUCATIONAL RESEARCH USE ONLY (RUO). MUST NOT BE USED IN DIAGNOSTIC OR OTHER MEDICAL APPLICATIONS. |
| Sensitivity: |
18.75pg/mL |
| Detection Limit: |
31.25~2000pg/mL |
| Short Description: |
This rat NFKB-p105 kit is a highly sensitive in-vitro chemiluminescent immunoassay for the measurement of trace amounts of analytes. |
| Storage Instruction: |
If unopened the kit may be stored at 2-8°C for up to 1 month. If the kit will not be used within 1 month, store the components separately, according to the component table in the manual. |
| Assay Time: |
3.5h |
| Detection: |
Chemiluminescence |
| Gene Symbol: |
Nfkb1Uniprot ID=NFKB1_RAT" |
| Specificity: |
This kit recognizes Rat NFKB-p105 in samples. No significant cross-reactivity or interference between Rat NFKB-p105 and analogues was observed. |
| Sample Type: |
Serum, plasma and other biological fluids |
| Post Translational Modifications | 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. A cotranslational processing allows the production of both p50 and p105 (Nuclear factor NF-kappa-B p105 subunit) from a single NFKB1 mRNA. 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. The GRR acts as a proteasomal 'stop signal', protecting the region upstream of the GRR from degradation and promoting generation of p50. It is unclear if limited proteasome degradation during cotranslational processing depends on ubiquitination. 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. Nuclear factor NF-kappa-B p105 subunit: 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). Phosphorylation at Ser-912 primes p105 for proteolytic processing in response to TNF-alpha stimulation. Phosphorylation at Ser-928, Ser-932 and Ser-937 are required for BTRC/BTRCP-mediated ubiquitination and proteolysis. Phosphorylation at Ser-932 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). Nuclear factor NF-kappa-B p105 subunit: Polyubiquitinated at multiple Lys residues in the C-terminus. Polyubiquitinated by the SCF(FBXW11) and SCF(BTRC) complexes following phosphorylation at Ser-928, Ser-932 and Ser-937, leading to its complete degradation. In contrast, polyubiquitination by the KPC complex following phosphorylation at Ser-932 leads to limited proteosomal processing and generation of the active NF-kappa-B p50 (Nuclear factor NF-kappa-B p50 subunit). S-nitrosylation of Cys-60 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. |
| 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. In a complex with MAP3K8, NFKB1/p105 represses MAP3K8-induced MAPK signaling.active MAP3K8 is released by proteasome-dependent degradation of NFKB1/p105. Nuclear factor NF-kappa-B p105 subunit: 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. Nuclear factor NF-kappa-B p50 subunit: 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. |
| Protein Name | Nuclear Factor Nf-Kappa-B P105 SubunitDna-Binding Factor Kbf1Ebp-1Nuclear Factor Of Kappa Light Polypeptide Gene Enhancer In B-Cells 1 Cleaved Into - Nuclear Factor Nf-Kappa-B P50 Subunit |
| Cellular Localisation | Nuclear Factor Nf-Kappa-B P105 Subunit: CytoplasmNuclear Factor Nf-Kappa-B P50 Subunit: NucleusCytoplasmAssociation With Nfkbia Inhibitor (I-Kappa-B)Promotes Its Retention In The Cytoplasm In An Inactive FormTranslocates Into The Nucleus Following Nfkbia Degradation |
| Alternative CLIA Names | Nuclear Factor Nf-Kappa-B P105 Subunit CLIA kitDna-Binding Factor Kbf1 CLIA kitEbp-1 CLIA kitNuclear Factor Of Kappa Light Polypeptide Gene Enhancer In B-Cells 1 Cleaved Into - Nuclear Factor Nf-Kappa-B P50 Subunit CLIA kitNfkb1 CLIA kit |
| Specificity | This kit recognizes Rat RANκ in samples. No significant cross-reactivity or interference between Rat RANκ and analogues was observed. |
| Reproducibility | Both intra-CV and inter-CV are |
Information sourced from Uniprot.org
| Item | Specifications | Storage |
| Micro CLIA Plate (Dismountable) | 96T: 8 wells ×12 strips strips | -20℃, 6 months |
| Reference Standard | 96T: 2 vials 48T: 1 vial | -20℃, 6 months |
| Concentrated Biotinylated Detection Ab (100×) | 96T: 1 vial, 120 μL 60 μL | -20℃, 6 months |
| Concentrated HRP Conjugate (100×) | 96T: 1 vial, 120 μL 60 μL | -20℃ (Protect from light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 2-8°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | 2-8°C, 6 months |
| HRP Conjugate Diluent | 1 vial, 14 mL | 2-8°C, 6 months |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | 2-8°C, 6 months |
| Substrate Reagent A | 1 vial, 5 mL | 2-8℃ (Protect from light) |
| Substrate Reagent B | 1 vial, 5 mL | 2-8℃ (Protect from light) |
| Plate Sealer | 5 pieces | |
| Manual | 1 copy | |
| Certificate of Analysis | 1 copy | |
| Sample Type | Range (%) | Average Recovery (%) |
| Serum(n=8) | 92-105 | 97 |
| EDTA plasma(n=8) | 92-108 | 98 |
| Cell culture media(n=8) | 97-111 | 105 |
| | Intra-assay Precision | Intra-assay Precision | Intra-assay Precision | Inter-assay Precision | Inter-assay Precision | Inter-assay Precision |
| Sample | 1.00 | 2.00 | 3.00 | 1.00 | 2.00 | 3.00 |
| n | 20.00 | 20.00 | 20.00 | 20.00 | 20.00 | 20.00 |
| Mean (pg/mL) | 106.37 | 186.04 | 892.11 | 104.72 | 188.76 | 855.40 |
| Standard deviation | 10.50 | 14.25 | 69.23 | 11.25 | 20.25 | 63.81 |
| CV (%) | 9.87 | 7.66 | 7.76 | 10.74 | 10.73 | 7.46 |
12 months for antibodies. 6 months for ELISA Kits. Please see website T&Cs for further guidance
