IKB alpha Antibody | Affinity Biosciences

$Fig. 4. Effects of stigmasterol treatment on inflammatory pathways in Aβ42 oligomers induced BV2 cells. Cells were pretreated with Aβ42 oligomers (1 μM) for 24 h, followed by treatment with stigmasterol (10, 20 μM) for 4 h. (A, B) Representative western blot analysis of AMPK signaling. GAPDH immunoreactivity was used as a loading control. (C-G) Representative western blot analysis of NF-κB signaling. The cytosolic fractions were prepared and analyzed with phosphorylated IκBα, total IκBα, and total NF-κB p65. The nuclear fractions were prepared and analyzed with total NF-κB p65. α-Tubulin immunoreactivity was used as a loading control in the cytosolic fraction, Histone H3 was used as the loading control in the nuclear fraction. (H-K) Representative western blot analysis of NLRP3 signaling, including NLRP3, Caspase-1, and Caspase-1, p20. GAPDH immunoreactivity was used as a loading control. Data were presented as the mean ± SEM from three independent experiments. One-way ANOVA with Tukey’s multiple comparison test revealed a difference between groups.$

Fig.

Figure 5 Effect of MALT1 mucosa-associated lymphoid tissue lymphoma translocation protein 1 on nuclear factor-κB signaling in proatherogenic VSMCs.

FIGURE 6 Effect of MLG on NF-κB/Nrf2/HO-1 signaling pathway in ethanol-induced gastric lesions mice.

Figure 5 Relative expression levels of NF-κB pathway-related proteins were measured via western blot assay.

Fig.

Product:	IKB alpha Antibody
Catalog:	AF5002
Description:	Rabbit polyclonal antibody to IKB alpha
Application:	WB IHC IF/ICC
Reactivity:	Human, Mouse, Rat
Prediction:	Pig, Bovine, Sheep, Rabbit, Dog, Chicken
Mol.Wt.:	39kDa; 36kD(Calculated).
Uniprot:	P25963
RRID:	AB_2834792

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

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MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human,Mouse,Rat

Prediction:

Pig(100%), Bovine(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(92%)

Clonality:

Polyclonal

Specificity:

IKB alpha Antibody detects endogenous levels of total IKB alpha.

RRID:

AB_2834792
Cite Format: Affinity Biosciences Cat# AF5002, RRID:AB_2834792.

Conjugate:

Unconjugated.

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Storage:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

Alias:

Fold/Unfold

I kappa B alpha; I-kappa-B-alpha; IkappaBalpha; IkB-alpha; IKBA; IKBA_HUMAN; IKBalpha; MAD 3; MAD3; Major histocompatibility complex enhancer-binding protein MAD3; NF kappa B inhibitor alpha; NF-kappa-B inhibitor alpha; NFKBI; NFKBIA; Nuclear factor of kappa light chain gene enhancer in B cells; Nuclear factor of kappa light polypeptide gene enhancer in B cells inhibitor alpha;

Immunogens

Immunogen:

Uniprot:

P25963(IKBA_HUMAN) >>Visit HPA database.

Gene(ID):

NFKBIA(4792)

Description:

NFKB1 (MIM 164011) or NFKB2 (MIM 164012) is bound to REL (MIM 164910), RELA (MIM 164014), or RELB (MIM 604758) to form the NFKB complex. The NFKB complex is inhibited by I-kappa-B proteins (NFKBIA or NFKBIB, MIM 604495), which inactivate NF-kappa-B by trapping it in the cytoplasm.

Sequence:

P25963 IKBA_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MFQAAERPQEWAMEGPRDGLKKERLLDDRHDSGLDSMKDEEYEQMVKELQEIRLEPQEVPRGSEPWKQQLTEDGDSFLHLAIIHEEKALTMEVIRQVKGDLAFLNFQNNLQQTPLHLAVITNQPEIAEALLGAGCDPELRDFRGNTPLHLACEQGCLASVGVLTQSCTTPHLHSILKATNYNGHTCLHLASIHGYLGIVELLVSLGADVNAQEPCNGRTALHLAVDLQNPDLVSLLLKCGADVNRVTYQGYSPYQLTWGRPSTRIQQQLGQLTLENLQMLPESEDEESYDTESEFTEFTEDELPYDDCVFGGQRLTL

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Pig

100

Bovine

100

Sheep

100

Dog

100

Rabbit

100

Chicken

Xenopus

Horse

Zebrafish

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P25963 As Substrate

P25963

Site	PTM Type	Enzyme	Source
	Ubiquitination		Uniprot
K21	Sumoylation		Uniprot
K21	Ubiquitination		Uniprot
K22	Sumoylation		Uniprot
K22	Ubiquitination		Uniprot
S32	Phosphorylation	P68400 (CSNK2A1) , O14965 (AURKA) , Q99558 (MAP3K14) , Q15418 (RPS6KA1) , Q14164 (IKBKE) , O00141 (SGK1) , P19525 (EIF2AK2) , P43250 (GRK6) , Q96KB5 (PBK) , O15111 (CHUK) , O14920 (IKBKB) , P51812 (RPS6KA3) , P34947 (GRK5) , Q9Y6K9 (IKBKG) , Q15349 (RPS6KA2)	Uniprot
S36	Phosphorylation	P68400 (CSNK2A1) , Q99558 (MAP3K14) , Q9UHD2 (TBK1) , P43250 (GRK6) , O15111 (CHUK) , Q15418 (RPS6KA1) , O14920 (IKBKB) , Q14164 (IKBKE) , O14965 (AURKA)	Uniprot
K38	Ubiquitination		Uniprot
Y42	Phosphorylation	P12931 (SRC) , P06213 (INSR) , P06239 (LCK) , P43405 (SYK)	Uniprot
K47	Ubiquitination		Uniprot
K67	Ubiquitination		Uniprot
K87	Ubiquitination		Uniprot
T90	Phosphorylation		Uniprot
K98	Ubiquitination		Uniprot
S166	Phosphorylation		Uniprot
K238	Ubiquitination		Uniprot
T273	Phosphorylation		Uniprot
S283	Phosphorylation	P68400 (CSNK2A1)	Uniprot
S288	Phosphorylation	P68400 (CSNK2A1)	Uniprot
T291	Phosphorylation	P68400 (CSNK2A1)	Uniprot
S293	Phosphorylation	P68400 (CSNK2A1)	Uniprot
T299	Phosphorylation	P68400 (CSNK2A1)	Uniprot
Y305	Phosphorylation	P00519 (ABL1) , A0A173G4P4 (Abl fusion)	Uniprot

Research Backgrounds

P25963_IKBA_HUMAN

Function:

Inhibits the activity of dimeric NF-kappa-B/REL complexes by trapping REL dimers in the cytoplasm through masking of their nuclear localization signals. On cellular stimulation by immune and proinflammatory responses, becomes phosphorylated promoting ubiquitination and degradation, enabling the dimeric RELA to translocate to the nucleus and activate transcription.

PTMs:

Phosphorylated; disables inhibition of NF-kappa-B DNA-binding activity. Phosphorylation at positions 32 and 36 is prerequisite to recognition by UBE2D3 leading to polyubiquitination and subsequent degradation.

Sumoylated; sumoylation requires the presence of the nuclear import signal. Sumoylation blocks ubiquitination and proteasome-mediated degradation of the protein thereby increasing the protein stability.

Monoubiquitinated at Lys-21 and/or Lys-22 by UBE2D3. Ubiquitin chain elongation is then performed by CDC34 in cooperation with the SCF(FBXW11) E3 ligase complex, building ubiquitin chains from the UBE2D3-primed NFKBIA-linked ubiquitin. The resulting polyubiquitination leads to protein degradation. Also ubiquitinated by SCF(BTRC) following stimulus-dependent phosphorylation at Ser-32 and Ser-36.

Deubiquitinated by porcine reproductive and respiratory syndrome virus Nsp2 protein, which thereby interferes with NFKBIA degradation and impairs subsequent NF-kappa-B activation.

Subcellular Location:

Cytoplasm. Nucleus.
Note: Shuttles between the nucleus and the cytoplasm by a nuclear localization signal (NLS) and a CRM1-dependent nuclear export.

Subunit Structure:

Interacts with RELA; the interaction requires the nuclear import signal. Interacts with NKIRAS1 and NKIRAS2. Part of a 70-90 kDa complex at least consisting of CHUK, IKBKB, NFKBIA, RELA, ELP1 and MAP3K14. Interacts with isoform 1 and isoform 2 of RWDD3; the interaction enhances sumoylation. Interacts (when phosphorylated at the 2 serine residues in the destruction motif D-S-G-X(2,3,4)-S) with BTRC. Associates with the SCF(BTRC) complex, composed of SKP1, CUL1 and BTRC; the association is mediated via interaction with BTRC. Part of a SCF(BTRC)-like complex lacking CUL1, which is associated with RELA; RELA interacts directly with NFKBIA. Interacts with PRMT2. Interacts with PRKACA in platelets; this interaction is disrupted by thrombin and collagen. Interacts with HIF1AN. Interacts with MEFV. Interacts with DDRGK1; positively regulates NFKBIA phosphorylation and degradation.

(Microbial infection) Interacts with HBV protein X.

Family&Domains:

Belongs to the NF-kappa-B inhibitor family.

Research Fields

· Cellular Processes > Cell growth and death > Apoptosis. (View pathway)

· Environmental Information Processing > Signal transduction > cAMP signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > NF-kappa B signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > TNF signaling pathway. (View pathway)

· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.

· Human Diseases > Infectious diseases: Bacterial > Epithelial cell signaling in Helicobacter pylori infection.

· Human Diseases > Infectious diseases: Bacterial > Shigellosis.

· Human Diseases > Infectious diseases: Bacterial > Legionellosis.

· Human Diseases > Infectious diseases: Parasitic > Leishmaniasis.

· Human Diseases > Infectious diseases: Parasitic > Chagas disease (American trypanosomiasis).

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Viral > Hepatitis C.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Measles.

· Human Diseases > Infectious diseases: Viral > Influenza A.

· Human Diseases > Infectious diseases: Viral > HTLV-I infection.

· Human Diseases > Infectious diseases: Viral > Herpes simplex infection.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer. (View pathway)

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Specific types > Prostate cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Chronic myeloid leukemia. (View pathway)

· Human Diseases > Cancers: Specific types > Small cell lung cancer. (View pathway)

· Organismal Systems > Immune system > Chemokine signaling pathway. (View pathway)

· Organismal Systems > Development > Osteoclast differentiation. (View pathway)

· Organismal Systems > Immune system > Toll-like receptor signaling pathway. (View pathway)

· Organismal Systems > Immune system > NOD-like receptor signaling pathway. (View pathway)

· Organismal Systems > Immune system > RIG-I-like receptor signaling pathway. (View pathway)

· Organismal Systems > Immune system > Cytosolic DNA-sensing pathway. (View pathway)

· Organismal Systems > Immune system > IL-17 signaling pathway. (View pathway)

· Organismal Systems > Immune system > Th1 and Th2 cell differentiation. (View pathway)

· Organismal Systems > Immune system > Th17 cell differentiation. (View pathway)

· Organismal Systems > Immune system > T cell receptor signaling pathway. (View pathway)

· Organismal Systems > Immune system > B cell receptor signaling pathway. (View pathway)

· Organismal Systems > Nervous system > Neurotrophin signaling pathway. (View pathway)

· Organismal Systems > Endocrine system > Adipocytokine signaling pathway.

· Organismal Systems > Endocrine system > Relaxin signaling pathway.

References

1). Kou L et al. Opsonized nanoparticles target and regulate macrophage polarization for osteoarthritis therapy: A trapping strategy. Journal of Controlled Release 2022 Jul;347:237-255. (PubMed: 35489544) [IF=10.8]

2). Li C et al. Oxyberberine, a novel gut microbiota-mediated metabolite of berberine, possesses superior anti-colitis effect: impact on intestinal epithelial barrier, gut microbiota profile and TLR4-MyD88-NF-κB pathway. PHARMACOLOGICAL RESEARCH 2019 Dec 18:104603 (PubMed: 31863867) [IF=9.3]

Application: WB Species: Mice Sample: colonic tissues

Fig. 6. Effect of OBB on the activation of TLR4-MyD88-NF-κB signaling pathway in DSS-induced colonic tissues. (A) Representative Western blotting images of TLR4, MyD88, cytoplasmic p65, nuclear p65, p-IκBα and IκBα. Changes in the relative protein expression levels of TLR4 (B), MyD88 (C), nuclear p65 (D), cytoplasmic p65 (E), and p-IκBα/IκBα ratio (F) were measured. Data are shown as the mean ± SEM (n = 3). # P < 0.05, ## P < 0.01 vs. Control group, * P < 0.05, ** P < 0.01 vs. DSS group.

3). Zhao C et al. Gut microbiota-mediated secondary bile acid alleviates Staphylococcus aureus-induced mastitis through the TGR5-cAMP-PKA-NF-κB/NLRP3 pathways in mice. npj Biofilms and Microbiomes 2023 Feb 8;9(1):8. (PubMed: 36755021) [IF=9.2]

4). Liu Y et al. Sophora japonica flowers and their main phytochemical, rutin, regulate chemically induced murine colitis in association with targeting the NF-κB signaling pathway and gut microbiota. Food Chemistry 2022 Jun 04;393:133395 (PubMed: 35691061) [IF=8.8]

5). Zhao C et al. Commensal cow Roseburia reduces gut-dysbiosis-induced mastitis through inhibiting bacterial translocation by producing butyrate in mice. Cell Reports 2022 Nov 22;41(8):111681. (PubMed: 36417859) [IF=8.8]

6). Li X et al. Semaphorin 7A interacts with nuclear factor NF-kappa-B p105 via integrin β1 and mediates inflammation. Cell Communication and Signaling 2023 Jan 30;21(1):24. (PubMed: 36717921) [IF=8.4]

Application: WB Species: Mouse Sample: livers

Fig. 2 The NF-κB pathway is activated in Sema7aR145W mouse liver and primary mouse hepatocytes. a–c Western blotting analysis of the relative protein levels of NF-κB p105, p- NF-κB p50/NF-κB p50, p- NF-κB p65/NF-κB p65 and p-IκB/IκB in the livers of wild-type (n = 4) and Sema7aR145W homozygous male mice (n = 5) and b, d in primary hepatocytes from wild-type (n = 3) and Sema7aR145W homozygous (n = 3) male mice. Phosphorylation levels were measured by the phosphor/total protein ratio. e Representative IF staining showing p-NF-κB p65 Ser529 (red) and DAPI (blue). Scale bars: 50 μm in liver sections from wild-type and Sema7aR145W homozygous male mice. f The relative levels of mRNA transcripts of the genes for NF-κB p105 in wild-type (n = 4) and Sema7aR145W homozygous male mice (n = 5) and g in primary hepatocytes from wild-type (n = 3) and Sema7aR145W homozygous (n = 3) male mice. The data were analysed by the independent-samples Student’s t test. * means p

7). Meng et al. S100A14 suppresses metastasis of nasopharyngeal carcinoma by inhibition of NF-kB signaling through degradation of IRAK1. Oncogene 2020 Jun 17. (PubMed: 32555330) [IF=8.0]

Application: WB Species: Human Sample: NPC cells

Fig 4.b NF-KB signaling makers in S100A14 overexpressing cells and S100A14 knocked-down cells were evaluated by immunoblotting.

8). Li CL et al. Comparison of anti-inflammatory effects of berberine, and its natural oxidative and reduced derivatives from Rhizoma Coptidis in vitro and in vivo. PHYTOMEDICINE 2019 Jan;52:272-283 (PubMed: 30599908) [IF=7.9]

9). Yuan L et al. Upregulation of UGT1A1 Expression by Ursolic Acid and Oleanolic Acid via the Inhibition of the PKC/NF-κB Signaling Pathway. Phytomedicine 2021 Aug 25;92:153726. (PubMed: 34536821) [IF=7.9]

Application: WB Species: Human Sample: HepG2 cells and Huh7 cells

Fig 2. Effects of PMA and LPS on UGT1A1 expression and NF-κB activity in HepG2 cells and Huh7 cells. HepG2 cells and Huh7 cells, respectively, were treated with PMA (100 nM) and/or LPS (10 μg/ml) for 24 h in the presence or absence of PDTC (20 μM). DMSO (0.1%) was used as the negative control. (A) Relative mRNA and (B) protein levels of UGT1A1 were quantified by qRT-PCR and western blot analyses, respectively. The experiments were normalized to GAPDH and compared to the negative control group. (C) Protein (cytoplasmic, nuclear or total) levels of p65, phospho-p65, IκBα, and phospho-IκBα were quantified by western blot analysis. Total-p65 was normalized to GAPDH, whereas the nucleus and cytoplasmic proteins were normalized to Lamin B1 and β-actin, respectively. All values were expressed as the mean ± S.E. of three independent experiments. (* p < 0.05, ** p < 0.01, *** p < 0.001).

10). Jie F et al. Stigmasterol attenuates inflammatory response of microglia via NF-κB and NLRP3 signaling by AMPK activation. Biomedicine & Pharmacotherapy 2022 Jun 27;153:113317. (PubMed: 35772378) [IF=7.5]

Application: WB Species: Mouse Sample: BV2 cells

Fig. 4. Effects of stigmasterol treatment on inflammatory pathways in Aβ42 oligomers induced BV2 cells. Cells were pretreated with Aβ42 oligomers (1 μM) for 24 h, followed by treatment with stigmasterol (10, 20 μM) for 4 h. (A, B) Representative western blot analysis of AMPK signaling. GAPDH immunoreactivity was used as a loading control. (C-G) Representative western blot analysis of NF-κB signaling. The cytosolic fractions were prepared and analyzed with phosphorylated IκBα, total IκBα, and total NF-κB p65. The nuclear fractions were prepared and analyzed with total NF-κB p65. α-Tubulin immunoreactivity was used as a loading control in the cytosolic fraction, Histone H3 was used as the loading control in the nuclear fraction. (H-K) Representative western blot analysis of NLRP3 signaling, including NLRP3, Caspase-1, and Caspase-1, p20. GAPDH immunoreactivity was used as a loading control. Data were presented as the mean ± SEM from three independent experiments. One-way ANOVA with Tukey’s multiple comparison test revealed a difference between groups.

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IKB alpha Antibody - #AF5002