Invitrogen

Phospho-ATF2 (Thr71) Polyclonal Antibody

高级验证

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated. 查看细节

查看其他45个ATF2抗体

说明书

Protocols

技术支持

说明书

Protocols

技术支持

Product Image Gallery

Phospho-ATF2 (Thr71) Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of Phospho-ATF-2 pThr71 showing staining in the nucleus of paraffin-embedded human breast carcinoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Anti- Phospho-ATF-2 pThr71 Polyclonal Antibody (Product # 44-295G) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Phospho-ATF2 (Thr71) Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts (30 µg lysate) of U-87 MG (Lane 1), U-87 MG treated with 25 µg/mL anisomycin for 30 minutes (Lane 2), K562 (Lane 3), K562 treated with 25 µg/mL anisomycin for 30 minutes (Lane 4). The blots were probed with Anti-Phospho-ATF-2 pThr71 Rabbit Polyclonal Antibody (Product # 44-295G, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (Product # G-21234, 1:5000 dilution). A ~ 54 kDa band corresponding to Phospho-ATF-2 pThr71 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Phospho-ATF2 (Thr71) Antibody — Altered expression of proteins upon cell treatment demonstrates antibody specificity. Western blot using Phospho-ATF2 (Thr71) Polyclonal Antibody (Product # 44-295G), shows increased levels of phospho-ATF2 (Thr71) in U-87MG and K562 cell lines upon Anisomycin treatment. {TM}

产品细节

Applications

Tested Dilution

Publications

免疫印迹 (WB)

1:1,000

免疫组化（石蜡） (IHC (P))

1:20

产品规格

种属反应

Human

宿主/亚型

Rabbit / IgG

分类

Polyclonal

类型

Antibody

抗原

Chemically synthesized phosphopeptide derived from a region of human ATF2 that contains threonine 71

偶联物

Unconjugated

形式

Liquid

纯化类型

Antigen affinity chromatography

保存液

Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/mL BSA

内含物

0.05% sodium azide

保存条件

-20°C

RRID

AB_2533624

产品详细信息

This antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non phosphorylated ATF2. The final product is generated by affinity chromatography using an ATF2-derived peptide phosphorylated at threonine 71. The peptide sequence is conserved in mouse, rat, chicken and frog. Suggested Western blot positive controls: NIH3T3 + anisomycin, or Jurkat + 10 µg/mL anisomycin for 60 minutes.

靶标信息

The transcription factor ATF-2 (also called CRE-BP1) binds to both AP-1 and CRE DNA response elements and is a member of the ATF/CREB family of leucine zipper proteins. ATF-2 interacts with a variety of viral oncoproteins and cellular tumor suppressors and is a target of the SAPK/JNK and p38 MAP kinase signaling pathways. Various forms of cellular stress, including genotoxic agents, inflammatory cytokines and UV irradiation, stimulate the transcriptional activity of ATF-2. Cellular stress activates ATF-2 by phosphorylation of Thr69 and Thr71. Both SAPK and p38 MAPK have been shown to phosphorylate ATF-2 at these sites in vitro and in cells transfected with ATF-2.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

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生物信息学

蛋白别名: Activating 2; Activating transcription factor 2; activating transcription factor 2 splice variant ATF2-var2; cAMP response element-binding protein CRE-BP1; cAMP responsive element binding protein 2, formerly; cAMP-dependent transcription factor ATF-2; CREB-2; Cyclic AMP-dependent transcription factor ATF-2; Cyclic AMP-responsive element-binding protein 2; HB16; histone acetyltransferase ATF2; MGC111558; OTTHUMP00000205345; OTTHUMP00000205347; OTTHUMP00000205348; OTTHUMP00000205350; OTTHUMP00000205352; OTTHUMP00000205353; OTTHUMP00000205354

基因别名: ATF2; CRE-BP1; CREB-2; CREB2; CREBP1; HB16; TREB7

UniProt ID: (Human) P15336

Entrez Gene ID: (Human) 1386

分子生物学功能: basic leucine zipper transcription factor