BMAL1 Antibody

Western Blot: BMAL1 Antibody [NB100-2288] - Oxidation causes accumulation of BMAL1. Head and neck cancer cells have different expression levels of the core clock protein BMAL1. Image collected and cropped by CiteAb from the following publication (//www.ncbi.nlm.nih.gov/pmc/articles/PMC5173143/) licensed under a CC-BY licence.

Immunohistochemistry: BMAL1 Antibody [NB100-2288] - Analysis of BMAL1 in mouse cerebral cortex (left: wt, right: Bmal1 KO) using anti-BMAL1 antibody. Image from verified customer review.

Immunohistochemistry: BMAL1 Antibody [NB100-2288] - Inhibition of mTOR signaling with Rapamycin rescues PTEN-driven accumulation of BMAL1A. Tissue samples and graphic show BMAL1 in K14cre PtenF/F mice and control (arrows). Note a significant amount of BMAL1 expression in PTEN mutant mice. K14cre PtenF/F mice and control littermates received Rapamycin or vehicle every other day for 15 days. Rapamycin treatment reduces the expression of nuclear BMAL1 in K14cre PtenF/F mice (arrowhead) (*** p0.05). Scale bars -30um. Image collected and cropped by CiteAb from the following publication (//www.ncbi.nlm.nih.gov/pmc/articles/PMC5173143/) licensed under a CC-BY licence.

Western Blot: BMAL1 Antibody [NB100-2288] - Oxidation causes accumulation of BMAL1. Western blot assay demonstrates time-dependent accumulation of BMAL1 upon oxidative stress in HNSCC cells (*p<0.05, **p<0.01). Image collected and cropped by CiteAb from the following publication (//www.ncbi.nlm.nih.gov/pmc/articles/PMC5173143/) licensed under a CC-BY licence.

Immunocytochemistry/Immunofluorescence: BMAL1 Antibody [NB100-2288] - Oxidation causes accumulation of BMAL1. Immunofluorescence assay depicts accumulation of ROS (green channel) and BMAL1 (red channel) upon oxidation. Image collected and cropped by CiteAb from the following publication (//www.ncbi.nlm.nih.gov/pmc/articles/PMC5173143/) licensed under a CC-BY licence.

Flow (Intracellular): BMAL1 Antibody [NB100-2288] - An intracellular stain was performed on HeLa cells with NB100-2288AF647(blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Knockout Validated: BMAL1 Antibody [NB100-2288] - CamiCre+; BMAL1fx/fx mice are forebrain/SCN-specific BMAL1 knockouts. (J) Immunohistochemistry for BMAL1 on SCN-containing coronal sections of Fx/Fx (n = 3), BKO (n = 3), and BMAL1-/- (KO, n = 1) mice sacrificed at ZT16. Captured with an 20x objective. F. CTX: frontal cortex, Cer: cerebellum. DOI://dx.doi.org/10.7554/eLife.04617.003Image collected and cropped by CiteAb from the following publication (//elifesciences.org/articles/04617) licensed under a CC-BY licence.

Flow Cytometry: BMAL1 Antibody [NB100-2288] - An intracellular stain was performed on Jurkat cells with NB100-2288 and a matched isotype control. Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody.

Western Blot: BMAL1 Antibody [NB100-2288] - Analysis of BMAL1 in A) MCF7, B) NIH/3T3, C) PC12.

Western Blot: BMAL1 Antibody [NB100-2288] - Analysis of MOP3 on 3T3/L1 cell lysates.

Immunocytochemistry/Immunofluorescence: BMAL1 Antibody [NB100-2288] - BMAL1 antibody was tested in HeLa cells at a 1:200 dilution against Dylight 488 (Green). Alpha tubulin and nuclei were counterstained against DyLight 568 (Red) and DAPI (Blue), respectively.

Immunohistochemistry: BMAL1 Antibody [NB100-2288] - Analysis of BMAL1 in mouse brain using DAB with hematoxylin counterstain.

Immunohistochemistry-Paraffin: BMAL1 Antibody [NB100-2288] - Analysis of a FFPE tissue section of mouse brain using 1:200 dilution of rabbit anti-BMAL1 antibody. The staining was developed using HRP labeled anti-rabbit IgG secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin. This BMAL1 antibody generated a specific nuclear staining in most of the cells and a relatively weaker cytoplasmic signal was also observed.

• Reactivity: Hu, Mu, Rt, Pm
• Applications: WB, ChIP, Flow, Flow-IC, ICC/IF, IHC, IHC-Fr, IHC-P, KO
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

BMAL1 Antibody Summary

• Immunogen: Bacterially expressed human BMAL1 (amino acids 392-626). [UniProt# O00327].
• Localization: Nuclear
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: ARNTL
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation
  • Flow (Intracellular)
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence 1:100
  • Immunohistochemistry 1:250
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:250
  • Knockout Validated
  • Western Blot 0.5 ug/mL - 2 ug/mL
• Application Notes: This BMAL1 antibody is useful in Immunocytochemistry/Immunofluorescence, Western blot, and IHC-paraffin embedded sections. Use in IHC-frozen sections was reported in the scientific literature (PMID: 23736292). In ICC/IF, primarily nuclear staining was observed with weak cytoplasmic staining in MCF7 cells. In Western Blot, a band was observed ~70 kDa. In IHC-P, staining was observed in the nuclei of mouse brain tissue. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. Use in IHC reported in scientific literature (PMID:33510438)
• Theoretical MW: 70 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  BMAL1 Overexpression Lysate

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID:32732906).

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: Tris-Glycine
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for BMAL1 Antibody

• ARNTL
• aryl hydrocarbon receptor nuclear translocator-like protein 1
• aryl hydrocarbon receptor nuclear translocator-like
• basic-helix-loop-helix-PAS orphan MOP3
• BHLHE5
• bHLHe5brain and muscle
• bHLH-PAS protein JAP3
• BMAL1
• BMAL1c
• BMAL1TIC
• Brain and muscle ARNT-like 1
• Class E basic helix-loop-helix protein 5
• JAP3
• Member of PAS protein 3
• member of PAS superfamily 3
• MOP3
• MOP3BMAL1c
• PAS domain-containing protein 3
• PASD3
• PASD3MGC47515
• TIC

Background

BMAL1 (brain and muscle ARNT-like 1; ARNTL) is an essential core component in circadian clock machinery which is regulatory mechanism for circadian rhythms. Circadian clock genes include three period proteins (PER1, PER2 and PER3), two cryptochromes (CRY1 and CRY2), CLOCK, NPAS2 and BMAL proteins, wherein BMAL1 has a potential to heterodimerize with CLOCK or NPAS2 genes; and this neocomplex drives transcription from E-box elements (5'-CACGTG-3') found in circadian-responsive genes's promoters. PER/CRY proteins negatively regulate CLOCK/BMAL1 dimer-mediated transcription, thereby forming the feedback loop that regulates the timing of clock gene transcription. BMAL1 also associates with GNB2L1/RACK1 and PRKCA in a nuclear complex, whertein GNB2L1 and PRKCA are recruited to the complex in a circadian manner. BMAL1 undergoes acetylation (Lys-538), phosphorylation and sumoylation (Lys-259) upon dimerization with CLOCK and acetylation facilitates CRY1-mediated repression. CLOCK-BMAL1 double mutations within PAS domains leads to syngernistic desensitization to high levels of CRY on repression of CLOCK-BMAL1 transcriptional activity of PER1 and, disrupt circadian rhythmicity. Clock genes functions primarily as tumor suppressors and their abbarant expression is observed in malignant pleural mesothelioma, colorectal and breast cancer.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for BMAL1 Antibody (NB100-2288)(34)

Publications using NB100-2288

• de Zavalia N, Schoettner K, Goldsmith JA Et al. Bmal1 in the striatum influences alcohol intake in a sexually dimorphic manner Communications biology Oct 26 2021 [PMID: 34702951] (WB, Mouse)
• Gegnaw ST, Sandu C, Mendoza J Et al. Dark-adapted light response in mice is regulated by a circadian clock located in rod photoreceptors Experimental eye research Oct 22 2021 [PMID: 34695438] (IHC, Mouse)
• Trebucq L, Cardama G, Lorenzano Menna P Et Al. Timing of Novel Drug 1A-116 to Circadian Rhythms Improves Therapeutic Effects against Glioblastoma Pharmaceutics Jul 16 2021 [PMID: 34371781] (WB)
• Huang W, Zheng X, Yang M et al. PER2-mediated ameloblast differentiation via PPAR gamma/AKT1/beta-catenin axis International journal of oral science May 19 2021 [PMID: 34011974]
• Kim H, Wei J, Song Z et al. Regulation of Hepatic Circadian Metabolism by the E3 ubiquitin ligase HRD1-controlled CREBH/PPARalpha Transcriptional Program Molecular metabolism Feb 13 2021 [PMID: 33592335] (WB, Mouse)
• Kim B, Kim J, Chun M, et al. Multiplexed CRISPR-Cas9 system in a single adeno-associated virus to simultaneously knock out redundant clock genes Scientific reports Jan 28 2021 [PMID: 33510438] (WB, IHC, Mouse)
• Wang XL, Wolff SEC, Korpel N et al. Deficiency of the Circadian Clock Gene Bmal1 Reduces Microglial Immunometabolism Frontiers in immunology Dec 8 2020 [PMID: 33363534] (WB, Mouse)
• Greco CM, Cervantes M, Fustin JM et al. S-adenosyl-l-homocysteine hydrolase links methionine metabolism to the circadian clock and chromatin remodeling Science advances Dec 1 2020 [PMID: 33328229] (ICC/IF, Mouse)
• Sominsky L, Dangel T, Malik S et al. Microglial ablation in rats disrupts the circadian system Nat Commun Nov 13 2020 [PMID: 33200466] (ICC/IF, WB, Human)
• Kim ER, Xu Y, Cassidy RM et al. Paraventricular hypothalamus mediates diurnal rhythm of metabolism Nat Commun Jul 30 2020 [PMID: 32732906] (IHC, Mouse)
• Hasegawa S, Fukushima H, Hosoda H et al. Hippocampal clock regulates memory retrieval via Dopamine and PKA-induced GluA1 phosphorylation Nat Commun Dec 18 2019 [PMID: 31852900] (WB, Mouse)
• Kim B, Kim J, Chun M et al. Single adeno-associated virus-based multiplexed CRISPR-Cas9 system to nullify core components of the mammalian molecular clock bioRxiv Jul 4 2020 (WB, Mouse)
• Dan H, Ruan T, Sampogna RV Circadian Clock Regulation of Developmental Time in the Kidney Cell Rep May 19 2020 [PMID: 32433970] (ICC/IF, Mouse)
• Petkau N TIP60 acetylation of BMAL1 links positive and negative arms of the molecular circadian clock Thesis (WB, ChIP, Mouse)
• Yuan P, Yang T, Mu J et al. Circadian clock gene NPAS2 promotes reprogramming of glucose metabolism in hepatocellular carcinoma cells Cancer Lett. Nov 22 2019 [PMID: 31765736] (ChIP, Human)
• Hannibal J, Norn THB, Georg B et al. Spatiotemporal expression pattern of PERIOD1 and PERIOD 2 in the mouse SCN is dependent on VIP receptor2 signaling Eur. J. Neurosci. Jun 18 2019 [PMID: 31211910]
• Luo Q, Xiao Y, Alex A et al. The Diurnal Rhythm of Insulin Receptor Substrate-1 (IRS-1) and Kir4.1 in Diabetes: Implications for a Clock Gene Bmal1 Invest. Ophthalmol. Vis. Sci. May 1 2019 [PMID: 31042800] (WB, Human, Rat)
• Cedernaes J, Huang W, Ramsey KM et al. Transcriptional Basis for Rhythmic Control of Hunger and Metabolism within the AgRP Neuron Cell Metab. Feb 18 2019 [PMID: 30827863] (IHC, Mouse)
• Zhu M, Tang H, Tang X et al. BMAL1 suppresses ROS-induced endothelial-to-mesenchymal transition and atherosclerosis plaque progression via BMP signaling. Am J Transl Res. Oct 15 2018 [PMID: 30416657] (WB, Human)
• Lananna B V, Nadarajah C J et al. Cell-Autonomous Regulation of Astrocyte Activation by the Circadian Clock Protein BMAL1. Cell Rep Feb 10 2018 [PMID: 30282019] (IHC, Mouse)
• Das V, Kc R, Li X, Varma D. Pharmacological targeting of the mammalian clock reveals a novel analgesic for osteoarthritis-induced pain. Gene. 2018 Feb 20 [PMID: 29474860] (Mouse)
• Kress GJ, Liao F, Dimitry J et al. Regulation of amyloid-B dynamics and pathology by the circadian clock. J. Exp. Med. Apr 2 2018 [PMID: 29382695] (ICC/IF, Mouse)
• Bostrom J, Sramkova Z, Salasova A et al. Comparative cell cycle transcriptomics reveals synchronization of developmental transcription factor networks in cancer cells. PLoS ONE 2017 Dec 11 [PMID: 29228002] (ICC/IF, Human)
• Lai B, Zou J, Lin Z et al. Haploinsufficiency of hnRNP U Changes Activity Pattern and Metabolic Rhythms Am. J. Pathol. 2017 Nov 09 [PMID: 29128567] (Mouse)
• Zagni C, Almeida L O et al. PTEN Mediates Activation of Core Clock Protein BMAL1 and Accumulation of Epidermal Stem Cells. Stem Cell Reports Nov 7 2017 [PMID: 28602615] (IHC, Human)
• Frederick A, Goldsmith J, de Zavalia N, Amir S. Mapping the co-localization of the circadian proteins PER2 and BMAL1 with enkephalin and substance P throughout the rodent forebrain. PLoS ONE. Apr 19 2017 [PMID: 28423013] (ICC/IF, Rat)
• Dumbell R, Leliavski A, Matveeva O et al. Dissociation of Molecular and Endocrine Circadian Rhythms in Male Mice Lacking Bmal1 in the Adrenal Cortex. Endocrinology 2016 Nov [PMID: 27690690] (Mouse)
• Matsumoto CS, Almeida LO, Guimaraes DM et al. PI3K-PTEN dysregulation leads to mTOR-driven upregulation of the core clock gene BMAL1 in normal and malignant epithelial cells. Oncotarget. Jul 5 2016 [PMID: 27285754] (ICC/IF, WB, Human)
• Perelis M, Marcheva B, Ramsey KM et al. Pancreatic B cell enhancers regulate rhythmic transcription of genes controlling insulin secretion. Science. 2015 Nov 06 [PMID: 26542580] (IHC-P, Mouse)
• Izumo M, Pejchal M, Schook AC et al. Differential effects of light and feeding on circadian organization of peripheral clocks in a forebrain Bmal1 mutant Elife. 2014 Dec 18 [PMID: 25525750] (WB, Mouse)
• Leliavski A, Shostak A, Husse J, Oster H. Impaired glucocorticoid production and response to stress in Arntl-deficient male mice. Endocrinology. 2014 Jan 01 [PMID: 24189141] (IHC-P)
• Obi-Ioka Y, Ushijima K, Kusama M et al. Involvement of Wee1 in the circadian rhythm dependent intestinal damage induced by docetaxel. J Pharmacol Exp Ther 2013 Jul 26 [PMID: 23892568] (ChIP, Mouse)
• Chu A, Zhu L, Blum ID et al. Global but not gonadotrope-specific disruption of bmal1 abolishes the luteinizing hormone surge without affecting ovulation. Endocrinology 2013 Aug [PMID: 23736292] (IHC-Fr, ICC/IF, Mouse)
• Hayashida S, Kuramoto Y, Koyanagi S et al. Proxisome proliferator-activated receptor-alpha mediates high-fat, diet-enhanced daily oscillation of plasminogen activator inhibitor-1 activity in mice. Chronobiol Int 2010 Oct [PMID: 20969520] (WB, ChIP, Mouse)

Review for BMAL1 Antibody (NB100-2288) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Mouse.

reviewed by: Verified Customer IF Mouse 06/29/2015

Summary

• Application: Immunofluorescence
• Sample Tested: mouse brain
• Species: Mouse

Blocking

• Blocking Details: 5% goat serum in TBS+0.05% Triton X-100 x 30min.

Primary Anitbody

• Dilution Ratio: NB100-2288 1:1000 in TBS+0.05% Triton X-100 + 1% goat serum. Incubated overnight at 4C.

Secondary Antibody

• Secondary Description: Alexaflor 568-conjugated Goat anti-rabbit, 1 hr incubation at room temp, in TBS+0.05% Triton X-10 . Then washed 3 x 5min in TBS

Details

• Detection Notes: Nikon fluorescent microscope, 568nm. Bmal1 KO brain tissue used as negative control (shown)
• Fixation Details: Mouse perfused with PBS+heparin, brain submerged in 3% paraformaldehyde x 24hrs, then 30% sucrose at -20C x 48 hrs.
• Wash Description: TBS: 3 x 5min washes

Comments

• Comments: Excellent antibody for IHC. Does not stain Bmal1 KO tissue (as many other Bmal1 antibodies do).

Bioinformatics

• Gene Symbol: ARNTL