Anti-CREB antibody [LB9] (ab178322)
Key features and details
- Mouse monoclonal [LB9] to CREB
- Suitable for: ICC/IF, WB, IP, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human, African green monkey
- Isotype: IgG1
Overview
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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-Pmore 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 Jurkat, NIH/3T3, C2C12, MDA-MB-231, T98G, 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.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 99% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
LB9 -
Isotype
IgG1 -
Research areas
Associated products
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Assay kits
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
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ICC/IF |
1/500.
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WB | (1) |
1/500. Detects a band of approximately 43 kDa (predicted molecular weight: 36 kDa).
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IP |
Use at an assay dependent concentration.
Suggested use 2 µg. |
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IHC-P |
1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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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. |
Target
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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
modificationsStimulated 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
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Database links
- Entrez Gene: 1385 Human
- Entrez Gene: 12912 Mouse
- Entrez Gene: 81646 Rat
- Omim: 123810 Human
- SwissProt: P16220 Human
- SwissProt: Q01147 Mouse
- SwissProt: P15337 Rat
- Unigene: 516646 Human
see all -
Alternative names
- Active transcription factor CREB antibody
- cAMP response element binding protein 1 antibody
- cAMP response element binding protein antibody
see all
Images
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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 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : K-562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate
Lane 4 : Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 5 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 6 : C2C12 whole cell lysate
Lane 7 : U-2 OS (Human bone osteosarcoma epithelial cell line) whole cell lysate
Lane 8 : A549 (Human lung carcinoma cell line) whole cell lysate
Lane 9 : MDA-MB-231 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 10 : T98G whole cell lysate
Lysates/proteins at 30 µg per lane.
Predicted band size: 36 kDa
Observed band size: 43 kDa why is the actual band size different from the predicted? -
Immunohistochemical analysis of deparaffinized Human tonsil tissue labeling CREB with ab178322 at 1/100 dilution followed by DAB staining.
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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.
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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 bladder cancer (left) and rectal cancer (right) tissues labeling CREB with ab178322 at 1/200 dilution followed by DAB staining.
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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.
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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.
Protocols
Datasheets and documents
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Datasheet download
References (15)
ab178322 has been referenced in 15 publications.
- Wei X et al. Silencing of Rab23 by siRNA inhibits ultraviolet B-induced melanogenesis via downregulation of PKA/CREB/MITF. Exp Dermatol 31:1253-1263 (2022). PubMed: 35514241
- Sohn E et al. Ficus erecta Thunb Leaves Alleviate Memory Loss Induced by Scopolamine in Mice via Regulation of Oxidative Stress and Cholinergic System. Mol Neurobiol 58:3665-3676 (2021). PubMed: 33797061
- Li T et al. Sevoflurane protects against ischemia-reperfusion injury in mice after total knee arthroplasty via facilitating RASD1-mediated protein kinase A pathway activation. Aging (Albany NY) 13:13333-13348 (2021). PubMed: 33982674
- Kong T et al. CD44 Promotes PD-L1 Expression and Its Tumor-Intrinsic Function in Breast and Lung Cancers. Cancer Res 80:444-457 (2020). PubMed: 31722999
- Misrani A et al. Citalopram prevents sleep-deprivation-induced reduction in CaMKII-CREB-BDNF signaling in mouse prefrontal cortex. Brain Res Bull 155:11-18 (2020). PubMed: 31743748