BMAL1 Antibody

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

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-Paraffin: BMAL1 Antibody [NB100-2288] - IHC analysis of a formalin fixed paraffin embedded 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.

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

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

Flow Cytometry: BMAL1 Antibody [NB100-2288] - An intracellular stain was performed on Jurkat cells with BMAL1 Antibody 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, Dylight 550 (SA5-10033, Thermo Fisher).

Chromatin Immunoprecipitation: 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.

Flow (Intracellular): BMAL1 Antibody [NB100-2288] - An intracellular stain was performed on HeLa cells with BMAL1 Antibody 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.

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

BMAL1 Antibody Summary

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

Applications/Dilutions

• Dilutions:
  • Western Blot 0.5 ug/mL - 2 ug/mL
  • Chromatin Immunoprecipitation
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence 1:100
  • Immunohistochemistry 1:250
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:250
  • Flow (Intracellular)
• 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.
• 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 Lysate (NBL1-07722)

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

• 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
• BMAL1TIC
• Brain and muscle ARNT-like 1
• Class E basic helix-loop-helix protein 5
• Member of PAS protein 3
• member of PAS superfamily 3
• MOP3BMAL1c
• PAS domain-containing protein 3
• PASD3MGC47515

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)(12)

Publications using NB100-2288

• 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)
• 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)
• 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)
• 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)
• Lauzier A, Charbonneau M, Harper K et al. Formation of invadopodia-like structures by synovial cells promotes cartilage breakdown in collagen-induced arthritis: Involvement of the protein tyrosine kinase Src. Arthritis Rheum;63(6):1591-1602. 2011 Jun. [PMID: 21337539] (ICC/IF, Rat)
• 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: Anonymous 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