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核因子κB抑制蛋白α抗體

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中文名稱 核因子κB抑制蛋白α抗體
別    名 NFKBIA; Inhibitor of KB alpha; I kappa B alpha; I(Kappa)B(alpha); IkappaBalpha; IKBA; IKBalpha; MAD 3; MAD3; Major histocompatibility complex enhancer binding protein MAD3; 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; IKBA_HUMAN. 

 

研究領(lǐng)域 腫瘤  免疫學(xué)  信號(hào)轉(zhuǎn)導(dǎo)  轉(zhuǎn)錄調(diào)節(jié)因子  激酶和磷酸酶  
抗體來(lái)源 Rabbit
克隆類型 Polyclonal
交叉反應(yīng) Human, Mouse, Rat, 
產(chǎn)品應(yīng)用 WB=1:500-2000 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 Flow-Cyt=1μg/Test IF=1:100-500 (石蠟切片需做抗原修復(fù))
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.
分 子 量 36kDa
細(xì)胞定位 細(xì)胞核 細(xì)胞漿 
性    狀 Liquid
濃    度 1mg/ml
免 疫 原 KLH conjugated synthetic peptide derived from human IKB alpha:1-120/314 
亞    型 IgG
純化方法 affinity purified by Protein A
儲(chǔ) 存 液 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
保存條件 Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
PubMed PubMed
產(chǎn)品介紹 Three major forms of IKB like molecules have been identified and each is characterised by multiple copies of ankyrin repeats. IKB alpha and IKB beta appear to be the major regulatory forms of IKB in most cells. These proteins interact with p65 or cRel containing forms of NFkB and block nuclear import by masking the nuclear localisation sequences of NFkB. The activation of NFkB involves the inducible phosphorylation and subsequent degradation of IKB. Immunoblotting easily detects the hyperphosphorylated forms of IKB alpha, but not phosphorylated IKB beta. Interestingly, IKB alpha and IKB beta mediate different NFkB responses. IkB alpha appears to control more transient activation of NFkB in response to an inducer, while IKB beta controls a persistent response. Bcl3 interacts with p50 and p52 containing forms of NFkB, but rather than being an inhibitor it appears to function to stimulate transcription. The degradation of IKB is confirmed by immunoblotting.

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.

Subunit:
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, IKBKAP and MAP3K14. Interacts with HBV protein X. Interacts with 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.

Subcellular Location:
Cytoplasm. Nucleus.

Tissue Specificity:
Highly expressed in lymph node, thymus followed by liver, brain, muscle, kidney, gastrointestinal and reproductive tract.

Post-translational modifications:
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.
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.

DISEASE:
Defects in NFKBIA are the cause of ectodermal dysplasia anhidrotic with T-cell immunodeficiency autosomal dominant (ADEDAID) [MIM:612132]. Ectodermal dysplasia defines a heterogeneous group of disorders due to abnormal development of two or more ectodermal structures. ADEDAID is an ectodermal dysplasia associated with decreased production of pro-inflammatory cytokines and certain interferons, rendering patients susceptible to infection.

Similarity:
Belongs to the NF-kappa-B inhibitor family.
Contains 5 ANK repeats.

SWISS:
Q9Z1E3

Gene ID:
4792

Database links:

Entrez Gene: 4792 Human

Entrez Gene: 18035 Mouse

Entrez Gene: 25493 Rat

Omim: 164008 Human

SwissProt: P25963 Human

SwissProt: Q9Z1E3 Mouse

SwissProt: Q63746 Rat

Unigene: 81328 Human

Unigene: 170515 Mouse

Unigene: 12550 Rat



Important Note:
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.

轉(zhuǎn)錄調(diào)節(jié)因子(Transcriptin Regulators)
產(chǎn)品圖片 Sample:
Thymus gland (Mouse) Lysate at 40 ug
Primary: Anti-IKB alpha (bs-1287R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 36 kD
Observed band size: 36 kD
Sample:
Spleen (Mouse) Lysate at 40 ug
Primary: Anti-IKB alpha (bs-1287R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 36 kD
Observed band size: 36 kD
Sample: Spleen (mouse) Lysate at 40 ug
Primary: Anti- IKB alpha (bs-1287R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 36 kD
Observed band size: 35 kD
Sample:
Lane 1: HepG2 (Human) Cell Lysate at 30 ug
Lane 2: Hela (Human) Cell Lysate at 30 ug
Lane 3: Siha (Human) Cell Lysate at 30 ug
Lane 4: U251 (Human) Cell Lysate at 30 ug
Lane 5: Spleen (Mouse) Lysate at 40 ug
Lane 6: Thymus (Mouse) Lysate at 40 ug
Lane 7: Thymus (Rat) Lysate at 40 ug
Lane 8: Lymph node (Mouse) Lysate at 40 ug
Lane 9: Lymph node (Rat) Lysate at 40 ug
Primary: Anti-IKB alpha (bs-1287R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 39 kD
Observed band size: 37 kD
Sample:
Stomach(Mouse) Lysate at 40 ug
Stomach(Rat) Lysate at 40 ug
Primary: Anti-IKB alpha (bs-1287R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 35 kD
Observed band size: 37 kD
Sample:
Lymph node (Mouse) Lysate at 40 ug
Thymus (Mouse) Lysate at 40 ug
Primary: Anti- IKB alpha (bs-1287R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 36 kD
Observed band size: 36 kD
Sample:
HepG2 (Human) Cell Lysate at 30 ug
Primary: Anti-IKB alpha (bs-1287R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 36 kD
Observed band size: 36 kD
Sample:
Hela(Human) Cell Lysate at 30 ug
Primary: Anti-IKB alpha (bs-1287R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 36 kD
Observed band size: 36 kD
Sample:
skin (Mouse) Lysate at 40 ug
Primary: Anti-IKB alpha (Bs- 1287R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 36 kD
Observed band size: 35 kD
Sample: Lung (mouse) Lysate at 40 ug
Primary: Anti- IKB alpha (bs-1287R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 36 kD
Observed band size: 36 kD
Paraformaldehyde-fixed, paraffin embedded (mouse lung); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (IKB alpha) Polyclonal Antibody, Unconjugated (bs-1287R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (IKB alpha) Polyclonal Antibody, Unconjugated (bs-1287R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Tissue/cell: rat kidney tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-IKB alpha Polyclonal Antibody, Unconjugated(bs-1287R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-IKB alpha antibody (bs-1287R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF647
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.Blank control (blue line): Jurkat (blue).
Primary Antibody (green line): Rabbit Anti-IKB alpha antibody (bs-1287R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 1μg /test.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min) , then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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