Antibody Panel to CRE Binding Proteins - Product Review 18

Introduction

The bZIP superfamily of eukaryotic DNA-binding transcription factors groups together proteins that contain a basic region mediating sequence - specific DNA-binding followed by a leucine zipper required for dimerization. bZIP domains usually bind a pallindromic 6 nucleotide site, but the specificity can be altered by interaction with accessory factor.

Several structure of bZIP have been solved. The basic region and the leucine zipper form a contiguous alpha helice where the four hydrophobic residues of the leucine zipper are oriented on one side. This conformation allows dimerization in parallel and it bends the helices so that the newly functional dimer forms a flexible fork where the basic domains, at the N-terminal open end, can then interact with DNA. The two leucine zipper are therefore oriented perpendicular to the DNA.

Within this quite large superfamily the mammalian cAMP response element (CRE) binding proteins form a subfamily. This family contains CREB, CREM, ATF1-7 and LRF1.

They are members of the Ser/Thr protein kinase super family and the cdc2/cdkx subfamily.

By definition, all Cdks share the feature that their enzymatic activation requires the binding of a cyclin regulatory subunit. In most cases, full activation also requires phosphorylation of a threonine near the kinase active site. Ckds are turned off by the binding of the CIP or INK4 proteins.

The cAMP response element (CRE) is an octanucleotide motif (TGACGTCA) that mediates diverse transcriptionally regulatory effects. It was first identified as an inducible enhancer of genes that can be transcribed in response to increased cAMP levels. Some growth control genes such as FOS have CRE in their transcriptional regulatory region and their expression is induced by increase in the intracellular cAMP levels. By cDNA cloning, multiple CRE-binding proteins have been identified. CREB1, originally called simply CREB, was isolated by Gonzalez et al. (1989); a gene called CREB2, or CREBP1, but officially designated ATF2 (activating transcription factor-2) was cloned by Maekawa et al. (1989). All of the CRE-binding proteins have the leucine zipper structure linked to a cluster of basic amino acids in their DNA-binding domain. The regulatory element TGACGTCA is found upstream of a number of viral and cellular genes. This element has been demonstrated to mediate cyclic AMP induction of cellular genes and activation of viral genes. The CREB or ATF proteins bind to this motif and mediate activation by cAMP and the adenovirus E1A protein. Activity of members of the CRE binding protein family is modulated by phosphorylation.

Antibody Tools for Detection of CRE Binding Proteins

Acris Antibodies offers a range of antibodies for detection of CRE Binding Proteins including CREB, phosphorylated CREB, and a full range of ATF antibodies (ATF1-7).

Picture Gallery

Detection of ATF-2 phosphorylation in HeLa cell lysate with anti-phospho ATF-2 mAb (Cat. #SM7006). Lane A. Untreated cell lysate; Lane B. HeLa cells treated with TNF for 1 hr Western blot analysis of ATF6 on 15 µg of (A) Jurkat and (B) NIH3T3 cell lysates using Cat. # SM7007P. A protein band of approximately 90 kD was detected Western blot analysis of CREB in HeLa cell lysate with anti-CREB mAb (Cat. #. SM7017P). A protein band of approximate molecular weight of 43 kDa was detected

Fig.1: Detection of ATF-2 phosphorylation in HeLa cell lysate with anti-phospho ATF-2 mAb (Cat.no. SM7006P). Lane A. Untreated cell lysate; Lane B. HeLa cells treated with TNF for 1 hr.

Fig.2: Western blot analysis of ATF6 on 15 µg of (A) Jurkat and (B) NIH3T3 cell lysates using Cat.no. SM7007P. A protein band of approximately 90 kD was detected.

Fig. 3: Western blot analysis of CREB in HeLa cell lysate with anti-CREB mAb (Cat.no. SM7017P). A protein band of approximate molecular weight of 43 kDa was detected.

Confocal images of double immunostaining with phospho CREB (Ser133) antibody RA18008 (green) and calbindin (red) in the hippocampus using the sections from control and 24 hours reperfusion following 15 minute transient cerebral ischemia in rats. Yellow represents overlay of red and green.

Fig 4: Confocal images of double immunostaining with phospho CREB (Ser133) antibody RA18008 (green) and calbindin (red) in the hippocampus using the sections from control and 24 hours reperfusion following 15 minute transient cerebral ischemia in rats. Yellow represents overlay of red and green.

Available Antibodies to CRE Binding Proteins / ATF

Anti- CREB1 (CREB), CREB2 (CREBP1, ATF2), CREB3 (LZIP), CREB3L1 (OASIS), CREB3L2 (BBF2H7),, CREB3L4 (CREB4, AIBZIP, JAL), CREB5, CREBL1 (ATF6-beta, Protein G13), CREBL2 , CREM (ICER), ATF1, ATF2 (CREB2), ATF3, ATF4, ATF5 (ATFX), ATF6 (ATF6-alpha), ATF7 (ATFA)antibody

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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  

ATF1 antibody

Western blot (WB) analysis of ATF1 antibody (Cat.-No.: AP06680PU-N) at 1/500 dilution Lane1:Hela whole cell lysate Lane2:sp2/0 whole cell lysate Lane3:H9C2 whole cell lysate Lane4:Mouse brain tissue lysate Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, P, WB
0.1 mg / €350.00
  Acris Antibodies GmbH

ATF1 antibody

Western Blot analysis of ATF1 expression in transfected 293T cell line (H00000466-T02) by ATF1 MaxPab polyclonal antibody.

Lane 1: ATF1 transfected lysate(29.20 KDa).
Lane 2: Non-transfected lysate. Mouse Serum Hu E(detection), WB
50 µl / €370.00
  Abnova Taiwan Corp.

ATF1 antibody

ATF1 MaxPab rabbit polyclonal antibody. Western Blot analysis of ATF1 expression in mouse brain. Rabbit Purified Hu, Ms, Rt E(detection), WB
0.1 mg / €380.00
  Abnova Taiwan Corp.

ATF1 antibody

ATF1 Mouse IgM 6G10 Ascites Hu E, WB
0.2 ml / €370.00
  Abnova Taiwan Corp.

ATF1 antibody

Immunoperoxidase of monoclonal antibody to ATF1 on formalin-fixed paraffin-embedded human placenta. [antibody concentration 3 ug/ml] Mouse IgG1 3E7 Purified Hu E, P, WB
0.1 mg / €370.00
  Abnova Taiwan Corp.

ATF1 antibody

Human 721_B; WB Suggested Anti-ATF1 Antibody Titration: 0.2-1 ug/ml. ELISA Titer: 1:62500. Positive Control: 721_B cell lysate. ATF1 is supported by BioGPS gene expression data to be expressed in 721_B.; ATF1 antibody - middle region (ARP31366_P050) in Human 721_B cells using Western Blot Rabbit Aff - Purified African clawed frog, Bov, Hu, Ms, Rt, Ze WB
50 µg / €350.00
  AVIVA Systems Biology

ATF1 antibody

Human Thymus; WB Suggested Anti-ATF1 Antibody Titration: 0.2-1 ug/ml. ELISA Titer: 1:312500. Positive Control: Human Thymus; ATF1 antibody - N-terminal region (ARP38573_P050) in Human Thymus cells using Western Blot Rabbit Aff - Purified African clawed frog, Bov, Can, Hu, Ms, Rt WB
50 µg / €350.00
  AVIVA Systems Biology

ATF1 antibody

Human Placenta; WB Suggested Anti-ATF1 Antibody Titration: 0.2-1 ug/ml. ELISA Titer: 1:12500. Positive Control: Human Placenta; ATF1 antibody - C-terminal region (ARP38574_P050) in Human Placenta cells using Western Blot Rabbit Aff - Purified African clawed frog, Bov, Hu, Ms, Por, Rt, Ze WB
50 µg / €350.00
  AVIVA Systems Biology

ATF1 pSer63 antibody

Immunohistochemistry: ATF1 [S63] Antibody (EP1591(2)Y) [NBP1-40539] - Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-specific anti-ATF-1 Rabbit IgG EP1591(2)Y Hu, Ms, Rt ICC/IF, P, WB
0.1 ml / €480.00
  Novus Biologicals Inc.

ATF2 pSer490/498 antibody

Figure 1. Western blot of Human melanoma cells incubated with varying doses of the radiomimetic drug NCS showing specific immuno-labeling of the ~74k ATF2 protein phosphorylated at Ser490 and Ser498. Rabbit IgG Aff - Purified Hu, Rt C, WB
0.1 ml / €380.00
  Acris Antibodies GmbH

ATF2 antibody

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using ATF-2 antibody. Rabbit Aff - Purified Hu, Ms ICC/IF, P, WB
0.1 ml / €270.00
  Acris Antibodies GmbH

ATF2 antibody

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using ATF-2 antibody. Rabbit Aff - Purified Hu, Ms ICC/IF, P, WB
50 µl / €230.00
  Acris Antibodies GmbH

ATF2 antibody

Immunohistochemical analysis of paraffin- embedded human breast carcinoma tissue using ATF-2 antibody. Rabbit Aff - Purified Hu, Ms ICC/IF, P, WB
0.1 ml / €270.00
  Acris Antibodies GmbH

ATF2 antibody

Immunohistochemical analysis of paraffin- embedded human breast carcinoma tissue using ATF-2 antibody. Rabbit Aff - Purified Hu, Ms ICC/IF, P, WB
50 µl / €230.00
  Acris Antibodies GmbH

ATF2 pThr69/51 + pThr71/53 antibody

ATF2 Rabbit Purified Hu, Ms, Rt IP, P, WB
0.1 mg / €460.00
  Acris Antibodies GmbH

ATF2 pSer112/94 antibody

ATF2 Rabbit Aff - Purified Hu, Ms, Rt E, IP, P, WB
0.1 mg / €350.00
  Acris Antibodies GmbH

ATF2 pSer62/44 antibody

ATF2 Rabbit Aff - Purified Hu, Ms, Rt E, IP, P, WB
0.1 mg / €350.00
  Acris Antibodies GmbH

ATF2 pThr69/51 antibody

ATF2 Rabbit Aff - Purified Hu, Ms, Rt E, IP, P, WB
0.1 mg / €350.00
  Acris Antibodies GmbH

ATF2 pThr71/53 antibody

Immunohistochemistry (IHC) analyzes of p-ATF2 (pThr71/53) antibody (Cat.-No.: AP01531PU-N) in paraffin-embedded human breast carcinoma tissue Rabbit Aff - Purified Hu, Ms, Rt IP, P, WB
0.1 mg / €350.00
  Acris Antibodies GmbH

ATF2 pThr73/55 antibody

Immunohistochemistry (IHC) analyzes of p-ATF2 (pThr73/55) antibody (Cat.-No.: AP01532PU-N) in paraffin-embedded human breast carcinoma tissue. Rabbit Aff - Purified Hu, Ms, Rt IP, P, WB
0.1 mg / €350.00
  Acris Antibodies GmbH

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