Overview

  • Product name

    Anti-CREB antibody [LB9]
    See all CREB primary antibodies
  • Description

    Mouse monoclonal [LB9] to CREB
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC/IF, WB, IP, IHC-P, ELISAmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, African green monkey
    Predicted to work with: Non human primates
  • Immunogen

    Recombinant full length protein corresponding to Human CREB aa 1-341.
    Sequence:

    MTMESGAENQ QSGDAAVTEA ENQQMTVQAQ PQIATLAQVS MPAAHATSSA PTVTLVQLPN GQTVQVHGVI QAAQPSVIQS PQVQTVQSSC KDLKRLFSGT QISTIAESED SQESVDSVTD SQKRREILSR RPSYRKILND LSSDAPGVPR IEEEKSEEET SAPAITTVTV PTPIYQTSSG QYIAITQGGA IQLANNGTDG VQGLQTLTMT NAAATQPGTT ILQYAQTTDG QQILVPSNQV VVQAASGDVQ TYQIRTAPTS TIAPGVVMAS SPALPTQPAE EAARKREVRL MKNREAAREC RRKKKEYVKC LENRVAVLEN QNKTLIEELK ALKDLYCHKS D


    Database link: P16220

  • Positive control

    • HeLa cells; Human bladder and rectal cancer tissues; Human tonsil tissue; Human HeLa, 293T, HepG2, A431, A549, K562, MCF7 and U2Os whole cell lysates; Mouse MEF, 3T3L1 and C2C12 whole cell lyastes; Rat NRK whole cell lysate; Monkey COS7 whole cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab178322 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/500.
WB 1/500. Detects a band of approximately 43 kDa (predicted molecular weight: 36 kDa).
IP Use at an assay dependent concentration.

Suggested use 2 μg.

IHC-P 1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ELISA Use a concentration of 0.1 - 1 µg/ml.

Target

  • Function

    This protein binds the cAMP response element (CRE), a sequence present in many viral and cellular promoters. CREB stimulates transcription on binding to the CRE. Transcription activation is enhanced by the TORC coactivators which act independently of Ser-133 phosphorylation. Implicated in synchronization of circadian rhythmicity.
  • Involvement in disease

    Defects in CREB1 may be a cause of angiomatoid fibrous histiocytoma (AFH) [MIM:612160]. A distinct variant of malignant fibrous histiocytoma that typically occurs in children and adolescents and is manifest by nodular subcutaneous growth. Characteristic microscopic features include lobulated sheets of histiocyte-like cells intimately associated with areas of hemorrhage and cystic pseudovascular spaces, as well as a striking cuffing of inflammatory cells, mimicking a lymph node metastasis. Note=A chromosomal aberration involving CREB1 is found in a patient with angiomatoid fibrous histiocytoma. Translocation t(2;22)(q33;q12) with CREB1 generates a EWSR1/CREB1 fusion gene that is most common genetic abnormality in this tumor type.
  • Sequence similarities

    Belongs to the bZIP family.
    Contains 1 bZIP domain.
    Contains 1 KID (kinase-inducible) domain.
  • Post-translational
    modifications

    Stimulated by phosphorylation. Phosphorylation of both Ser-133 and Ser-142 in the SCN regulates the activity of CREB and participates in circadian rhythm generation. Phosphorylation of Ser-133 allows CREBBP binding (By similarity). Phosphorylated upon DNA damage, probably by ATM or ATR.
    Sumoylated by SUMO1. Sumoylation on Lys-304, but not on Lys-285, is required for nuclear localization of this protein. Sumoylation is enhanced under hypoxia, promoting nuclear localization and stabilization.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • Active transcription factor CREB antibody
    • cAMP response element binding protein 1 antibody
    • cAMP response element binding protein antibody
    • cAMP responsive element binding protein 1 antibody
    • cAMP-responsive element-binding protein 1 antibody
    • CREB antibody
    • CREB-1 antibody
    • CREB1 antibody
    • CREB1_HUMAN antibody
    • Cyclic AMP-responsive element-binding protein 1 antibody
    • MGC9284 antibody
    • OTTHUMP00000163864 antibody
    • OTTHUMP00000163865 antibody
    • OTTHUMP00000206660 antibody
    • OTTHUMP00000206662 antibody
    • OTTHUMP00000206667 antibody
    • Transactivator protein antibody
    see all

Images

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: CREB knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: A431 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab178322 observed at 44 kDa. Red - loading control, ab18058, observed at 124 kDa.

    ab178322 was shown to recognize CREB when CREB knockout samples were used, along with additional cross-reactive bands. Wild-type and CREB knockout samples were subjected to SDS-PAGE.  Ab178322 and ab18058 (loading control to Vinculin) were diluted at 1/250 and 1/10,000 dilution respectively and incubated overnight at 4C. Blots were developed with IRDye® 800CW Goat anti-Rabbit IgG (H + L) and IRDye® 680 Goat anti-Mouse IgG (H + L) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-CREB antibody [LB9] (ab178322) at 1/500 dilution

    Lane 1 : HeLa whole cell lysate
    Lane 2 : 293T whole cell lysate
    Lane 3 : A431 whole cell lysate
    Lane 4 : A549 whole cell lysate
    Lane 5 : MEF whole cell lysate
    Lane 6 : MCF7 whole cell lysate
    Lane 7 : U2OS whole cell lysate
    Lane 8 : K562 whole cell lysate
    Lane 9 : COS7 whole cell lysate
    Lane 10 : 3T3L1 whole cell lysate
    Lane 11 : C2C12 whole cell lysate
    Lane 12 : NRK whole cell lysate

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : goat anti-mouse-HRP at 1/20000 dilution

    Developed using the ECL technique.

    Predicted band size: 36 kDa

  • Immunohistochemical analysis of deparaffinized Human tonsil tissue labeling CREB with ab178322 at 1/100 dilution followed by DAB staining.

  • Immunoprecipitation analysis of CREB was performed on untreated 293T cells. The antigen:antbody complex was formed by binding 500μg whole cell lysate with 2μg ab178322 and captured on 50μl Protein A/G Plus Agarose. 

    Lane 1: 293T lysate

    Lane 2: IP of 293T lysate

    WB detection used ab178322 at 1/1000 followed with a goat anti-mouse-HRP secondary antibody at 1/20,000.

  • Immunohistochemical analysis of deparaffinized Human bladder cancer (left) and rectal cancer (right) tissues labeling CREB with ab178322 at 1/200 dilution followed by DAB staining.

  • Immunofluorescent analysis of untreated HeLa cells (formalin-fixed, 0.1% Triton X-100 permeabilized) labeling CREB with ab178322 at 1/400 dilution (green) followed with DyLight 488 goat anti-mouse IgG secondary antibody at 1/400 dilution. Nuclei (blue) were stained with Hoechst 33342 dye.

  • Immunofluorescent analysis of untreated HeLa cells (formalin-fixed, 0.1% Triton X-100 permeabilized) labeling CREB with ab178322 at 1/500 dilution (green) followed with DyLight 488 goat anti-mouse IgG secondary antibody at 1/400 dilution. Nuclei (blue) were stained with Hoechst 33342 dye.

References

This product has been referenced in:

  • Tabassum S  et al. Jujuboside A prevents sleep loss-induced disturbance of hippocampal neuronal excitability and memory impairment in young APP/PS1 mice. Sci Rep 9:4512 (2019). Read more (PubMed: 30872728) »
  • Li X  et al. Cyclin-dependent kinase 5 regulates proliferation, migration, tyrosinase activity, and melanin production in B16-F10 melanoma cells via the essential regulator p-CREB. In Vitro Cell Dev Biol Anim 55:416-425 (2019). Read more (PubMed: 31069610) »
See all 5 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Snail Cell lysate - nuclear (Brain)
Gel Running Conditions
Reduced Denaturing (Pre-cast SDS-Gradient-Gel)
Loading amount
30 µg
Specification
Brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

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Verified customer

Submitted Aug 06 2019

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