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NF-kappaB Pathway Antibody Sampler Kit
NF-kappaB Pathway Antibody Sampler Kit
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NF-kappaB Pathway Antibody Sampler Kit

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    分子量：
    40 IkappaB-alpha. 65 p65/RelA. 85 IKKalpha. 87 IKKbeta.
    40 IkappaB-alpha. 65 p65/RelA. 85 IKKalpha. 87 IKKbeta.
    产品介绍
    产品信息
    抗原名称
    NF-kappaB Pathway
    来源纯化
    Monoclonal antibodies are produced by immunizing animals with recombinant human proteins or synthetic peptides.
    简单描述
    Antibody Sampler Kit for studying Mouse IgG in the research area.
    分子量
    40 IkappaB-alpha. 65 p65/RelA. 85 IKKalpha. 87 IKKbeta.
    应用
    目标/特异性

    Specificity/Sensitivity

    Phospho-IKKα/β, phospho-NF-κB p65, and phospho-IκBα antibodies recognize endogenous levels of IKKα/β, p65, and IκB-α, respectively, only when phosphorylated at the indicated residues. They do not cross-react with other family members at physiological levels. Total IKKα, IKKβ, p65, and IκBα antibodies recognize endogenous levels of their respective targets regardless of phosphorylation state and do not cross-react with other family members at physiological levels.

    背景
    背景
    The transcriptional nuclear factor κB (NF-κB)/Rel transcription factors are present in the cytosol in an inactive state, complexed with the inhibitory IκB proteins. Activation occurs via phosphorylation of IκBα at Ser32 and Ser36, resulting in the ubiquitin-mediated proteasome-dependent degradation of IκBα and the release and nuclear translocation of active NF-κB dimers. The regulation of IκBβ and IκBε is similar to that of IκBα, however, the phosphorylation and degradation of these proteins occurs with much slower kinetics. Phosphorylation of IκBβ occurs at Ser/Thr19 and Ser23, while IκBε can be phosphorylated at Ser18 and Ser22. The key regulatory step in this pathway involves activation of a high molecular weight IkappaB kinase (IKK) complex, consisting of three tightly associated IKK subunits. IKKα and IKKβ serve as the catalytic subunits of the kinase. Activation of IKK depends on phosphorylation at Ser177 and Ser181 in the activation loop of IKKβ (176 and 180 in IKKα). NF-κB-inducing kinase (NIK), TANK-binding kinase 1 (TBK1), and its homolog IKKε (IKKi), phosphorylate and activate IKKα and IKKβ.The NF-κB family of transcription factors is comprised of five proteins in mammals, p65/RelA, c-Rel, RelB, NF-κB1 (p105/p50) and NF-κB2 (p100/p52). p105 and p100 are proteolytically processed to produce p50 and p52, respectively. The 50 kDa active form is produced through proteolytic processing following IKK-mediated phosphorylation of p105 at multiple sites (Ser922, 924, 928 and 933), while p100's processing to p52 is induced by phosphorylation of Ser864 and Ser868. The p50 and p52 products form dimeric complexes with Rel proteins, which are then able to bind DNA and regulate transcription. Phosphorylation of p65/RelA at Ser276 by PKA C and MSK1 enhances transcriptional activity. p65 phosphorylation at Ser536 regulates activation, nuclear localization, protein-protein interactions, and transcriptional activity. PMA-induced NF-κB transcriptional activity is dependent on the region of p65 containing the potential phosphorylation sites Ser457, Thr458, Thr464 and Ser468. Phosphorylation of Ser468 by GSK-3β inhibits basal p65 activity. 1.Yamamoto, Y. and Gaynor, R.B. (2004) Trends Biochem. Sci. 29, 72-79. 2.Ghosh, S. and Karin, M. (2002) Cell 109, S81-S96. 3.Viatour, P. et al. (2005) Trends Biochem. Sci. 30, 43-52. 4.Ho, C. et al. (2016) PLOS One 10,1-22. 5.Beyaz S. et al. (2016) Nature 531, 53-58.
    翻译后修饰
    Unmodified

    参考图片

    Confocal immunofluorescent analysis of HeLa cells, untreated (left), or TNF-α-treated (#8902, 10 ng/ml for 15 min, right) using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

    Confocal immunofluorescent analysis of HeLa cells, untreated (left) and TNF-α treated (#8902 at 20 ng/ml for 20 min, right), using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® phalloidin 555 (red).

    Western blot analysis of extracts from THP-1 cells, differentiated with TPA (#9905, 80 nM for 24h) and treated with 1 μg/ml LPS for the indicated times, using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.

    Western blot analysis of extracts from THP-1 cells, differentiated with TPA (#9905, 80 nM for 24h) and treated with 1 μg/ml LPS for the indicated times, using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb (upper) and NF-κB p65 (C22B4) Rabbit mAb #4764 (lower).

    Western blot analysis of extracts from THP-1 cells, differentiated with TPA (#9905, 80 nM for 24h) and treated with 1 μg/ml LPS for the indicated times, using Phospho-IκBα (Ser32/36) (5A5) Mouse mAb #9246 (upper) and IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) (lower).

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.

    Immunohistochemical analysis of paraffin-embedded human gall bladder (chronic cholecystitis), using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.

    Immunohistochemical analysis of paraffin-embedded human lung (chronic bronchitis), using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.

    Flow cytometric analysis of HeLa cells, using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) (blue) compared to a nonspecifc negative control antibody (red).

    Western blot analysis of extracts from HeLa, NIH/3T3 and PC12 cells, using IkBα (L35A5) Mouse mAb (Amino-terminal Antigen).

    Western blot analysis of extracts from TNF-alpha and Calyculin A treated HeLa and NIH/3T3 cells, using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.

    Confocal immunofluorescent analysis of HT-1080 cells, untreated (left) or treated with hTNF-α #8902 (20 ng/ml, 20 min) (right), using NF-κB p65 (D14E12) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

    Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated or TNF-α treated (#2169, 20 ng/ml for 5 minutes), using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb (upper) or NF-κB p65 Antibody #3034 (lower).

    Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen).

    Immunohistochemical analysis of paraffin-embedded human renal adenocarcinoma, using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen).

    Immunohistochemical analysis of paraffin-embedded human leiomyoma, using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen).

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing cytoplasmic localization, using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.

    Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb in the presence of control peptide (left) or Phospho-IKK-alpha/beta (Ser176/180) Blocking Peptide #1023 (right).

    Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated or treated with TNF-α (#2169, 20 ng/ml) for 5 minutes, using Phospho-IκBα (Ser32) (14D4) Rabbit mAb (upper), or IκBα (44D4) Rabbit mAb #4812 (lower).

    Immunohistochemical analysis of frozen H1650 xenograft, showing cytoplasmic localization using Phospho-IKKα/β (Ser176/180)(16A6) Rabbit mAb.

    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.

    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.

    Flow cytometric analysis of HeLa cells, untreated (blue) or TNF-α-treated (green), using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb compared to a nonspecific negative control antibody (red).

    Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells treated with hTNF-α #8902 (30 ng/ml, 1 hr) and either 5 μl of NF-κB p65 (D14E12) XP® Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by Real-Time PCR using SimpleChIP® Human IκBα Promoter Primers #5552, human IL-8 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

    Western blot analysis of extracts from various cell lines using NF-κB p65 (D14E12) XP® Rabbit mAb.

    Flow cytometric analysis of HeLa cells using NF-κB p65 (D14E12) XP® Rabbit mAb (blue) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red).

    Immunohistochemical analysis of paraffin-embedded human chronic cholecystitis using NF-κB p65 (D14E12) XP® Rabbit mAb.

    Western blot analysis of extracts from various cell lines using NF-κB p65 (D14E12) XP® Rabbit mAb #8242.

    Western blot analysis of extracts from various cell lines using IKKβ (D30C6) Rabbit mAb.

    Western blot analysis of extracts from wild-type, IKKα (-/-), and IKKβ (-/-) mouse embryonic fibroblasts (MEFs) using IKKβ (D30C6) Rabbit mAb (upper) and GAPDH (14C10) Rabbit mAb #2118 (lower).

    Western blot analysis of extracts from NIH/3T3, HeLa and PC12 cells, using IKKα Antibody #2682.

    Western blot analysis of extracts from HeLa, NIH/3T3 and PC-12 cells using IIKKβ (2C8) Rabbit mAb #2370.

    Western blot analysis of extracts from TNF-alpha and Calyculin A treated HeLa and NIH/3T3 cells, using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb #2697.

    Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated or treated with TNF-α (#2169, 20 ng/ml) for 5 minutes, using Phospho-IκB-α (Ser32) (14D4) Rabbit mAb #2859 (upper), or IκBα (44D4) Rabbit mAb #4812 (lower).

    Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated or TNF-α treated (#2169, 20 ng/ml for 5 minutes), using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb #3033 (upper) or NF-κB p65 Antibody #3034 (lower).

    Western blot analysis of extracts from HeLa, NIH/3T3 and PC12 cells, using IκB-α (L35A5) Mouse mAb (Amino-terminal Antigen) #4814.

    Confocal immunofluorescent analysis of HCT 116 (high expression; left) and IGROV-1 (low expression; right) cells using IKKα (3G12) Mouse mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

    Western blot analysis of extracts from various cell lines using IKKα (3G12) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

    Flow cytometric analysis of HCT 116 cells using IKKα (3G12) Mouse mAb (blue) compared to concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (red). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.

    Flow cytometric analysis of THP-1 cells, untreated (blue) and with TPA and LPS (green) using IKK-α (Ser176/Ser180) phosphate Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (PE Conjugate) #8885 was used as a secondary antibody.

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    货号：
    9936T
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