Anticorps Recombinant de lapin anti-ATF4

ATF4 Recombinant Antibody for WB, IHC, IP, ELISA, ChIP-qPCR

Hôte / Isotype

Lapin / IgG

Réactivité testée

Humain, souris

Applications

WB, IHC, IP, ELISA, ChIP-qPCR

Conjugaison

Non conjugué

CloneNo.

5N3

N° de cat : 81798-1-RR

Synonymes

5N3, cAMP-dependent transcription factor ATF-4, CREB2, Cyclic AMP-responsive element-binding protein 2, Tax-responsive enhancer element-binding protein 67

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Galerie de données de validation

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  WB analysis of HEK-293 using 81798-1-RR

  Non-treated HEK-293 and MG132 treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 81798-1-RR (ATF4 antibody) at dilution of 1:15000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

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  WB analysis of A431 using 81798-1-RR

  A431 cells were subjected to SDS PAGE followed by western blot with 81798-1-RR (ATF4 antibody) at dilution of 1:15000 incubated at room temperature for 1.5 hours.

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  WB analysis of HeLa using 81798-1-RR

  Non-treated HeLa and MG132 treated HeLa cells were subjected to SDS PAGE followed by western blot with 81798-1-RR (ATF4 antibody) at dilution of 1:15000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

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  IP experiment of HEK-293 using 81798-1-RR

  IP result of anti-ATF4 (IP:81798-1-RR, 4ug; Detection:81798-1-RR 1:4000) with HEK-293 cells lysate 1440 ug.

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  IHC staining of human breast cancer using 81798-1-RR

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 81798-1-RR (ATF4 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 81798-1-RR

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 81798-1-RR (ATF4 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human stomach cancer using 81798-1-RR

  Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 81798-1-RR (ATF4 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human stomach cancer using 81798-1-RR

  Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 81798-1-RR (ATF4 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human colon cancer using 81798-1-RR

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 81798-1-RR (ATF4 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  ChIP experiment of NIH/3T3 using 81798-1-RR

  Chromatin was prepared from NIH/3T3 cells treated with Tunicamycin (2ug/mll) overnight. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 20 µg of cross-linked chromatin, 5 µg of ATF4 (81798-1-RR) or 5 ug of Normal Rabbit IgG (98136-1-RR), and 20 µl of Protein A Magarose Beads. The immunoprecipitated DNA was quantified by real-time PCR.

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  Affinity and Kinetic Characterization of 81798-1-RR

  Biolayer interferometry (BLl) kinetic assays of 81798-1-RR against Human ATF4 were performed. The affinity constant is 0.500 nM.

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Applications testées

• Résultats positifs en WB: cellules A431, MG132 treated HEK-293 cells, MG132 treated HeLa cells
• Résultats positifs en IP: cellules HEK-293,
• Résultats positifs en IHC: tissu de cancer du sein humain, tissu de cancer de l'estomac humain, tissu de cancer du côlon humain; il est suggéré de démasquer l'antigène avec un tampon de TE buffer pH 9.0; (*) À défaut, 'le démasquage de l'antigène peut être 'effectué avec un tampon citrate pH 6,0.
• Résultats positifs en ChIP-qPCR: Tunicamycin (2ug/mll) overnight NIH/3T3 cells,

Dilution recommandée

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprécipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochimie (IHC): IHC : 1:50-1:500
• CHImmunoprécipitation (IP)-QPCR: CHIP-QPCR : 1:10-1:100

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, check data in validation data gallery

Informations sur le produit

81798-1-RR cible ATF4 dans les applications de WB, IHC, IP, ELISA, ChIP-qPCR et montre une réactivité avec des échantillons Humain, souris

• Réactivité: Humain, souris
• Hôte / Isotype: Lapin / IgG
• Clonalité: Recombinant
• Type: Anticorps
• Immunogène: ATF4 Protéine recombinante Ag1279
• Nom complet: activating transcription factor 4 (tax-responsive enhancer element B67)
• Masse moléculaire calculée: 39 kDa
• Poids moléculaire observé: 50 kDa
• Numéro d’acquisition GenBank: BC022088
• Symbole du gène: ATF4
• Identification du gène (NCBI): 468
• Conjugaison: Non conjugué
• Forme: Liquide
• Méthode de purification: Purification par protéine A
• Tampon de stockage: PBS with 0.02% sodium azide and 50% glycerol
• Conditions de stockage: Stocker à -20°C. Stable pendant un an après l'expédition. L'aliquotage n'est pas nécessaire pour le stockage à -20^oC Les 20ul contiennent 0,1% de BSA.

Informations générales

ATF4 is a transcription factor, that accumulates predominantly in osteoblasts, where it regulates terminal osteoblast differentiation and bone formation[PMID: 19016586]. As a basic leucine-zipper (bZip) transcription factor, ATF4 can regulate amino acid metabolism, cellular redox state, and anti-stress responses. It also regulates age-related and diet-induced obesity and glucose homeostasis in mammals, and has conserved metabolic functions in flies[PMID: 19726872]. Due to its location at chromosome 22q13, a region linked to schizophrenia, ATF4 is considered as a positional candidate gene for schizophrenia[PMID: 18163433]. Otherwise, since ATF4 is induced by tumour microenvironmental factors, and regulates processes relevant to cancer progression, it might serve as a potential therapeutic target in cancer. Endogenous ATF4 protein has a molecular mass of 50kd. [PMID: 17726049]. ATF4 can bind DNA as a homodimer and as a heterodimer. ATF4 is ubiquitinated by SCF(BTRC) in response to mTORC1 signal, followed by proteasomal degradation and leading to down-regulate expression of SIRT4, so the molecular weight of ATF4 may be 70 kDa.

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