PA1-523

antibody from Invitrogen Antibodies

Targeting: ARNTL bHLHe5, BMAL1, JAP3, MOP3, PASD3

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-523

Provider

Invitrogen Antibodies

Product name

BMAL1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

PA1-523 detects BMAL1/aryl hydrocarbon nuclear translocator 3 (ARNT3) from hamster and mouse tissues as well as recombinant human BMAL1. PA1-523 has been sucessfully used in Western blot procedures. By Western blot, this antibody detects a 110 kDa protein which corresponds to the product of a hamster GST-BMAL1 fusion construct overexpressed in E. coli. This antibody detects a non-specific band in U87-MG cell lysates at ~105kDa and in NIH-3T3 cell lysates at ~135kDa. PA1-523 immunizing peptide corresponds to amino acid residues 582-594 from mouse BMAL1. This sequence is completely conserved between mouse, rat, guinea pig, and human BMAL1. PA1-523 immunizing peptide (Cat. # PEP-075) is available for use in neutralization and control experiments.

Reactivity

Human, Mouse, Rat, Hamster

Host

Rabbit

Isotype

IgG

Vial size

200 µL

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

Prolonged Administration of Melatonin Ameliorates Liver Phenotypes in Cholestatic Murine Model.

Ceci L, Chen L, Baiocchi L, Wu N, Kennedy L, Carpino G, Kyritsi K, Zhou T, Owen T, Kundu D, Sybenga A, Isidan A, Ekser B, Franchitto A, Onori P, Gaudio E, Mancinelli R, Francis H, Alpini G, Glaser S
Cellular and molecular gastroenterology and hepatology 2022;14(4):877-904

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Melatonin modulates daytime-dependent synaptic plasticity and learning efficiency.

Jilg A, Bechstein P, Saade A, Dick M, Li TX, Tosini G, Rami A, Zemmar A, Stehle JH
Journal of pineal research 2019 Apr;66(3):e12553

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Dysregulated circadian rhythm pathway in human osteoarthritis: NR1D1 and BMAL1 suppression alters TGF-β signaling in chondrocytes.

Akagi R, Akatsu Y, Fisch KM, Alvarez-Garcia O, Teramura T, Muramatsu Y, Saito M, Sasho T, Su AI, Lotz MK
Osteoarthritis and cartilage 2017 Jun;25(6):943-951

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The transcription factor Runx2 is under circadian control in the suprachiasmatic nucleus and functions in the control of rhythmic behavior.

Reale ME, Webb IC, Wang X, Baltazar RM, Coolen LM, Lehman MN
PloS one 2013;8(1):e54317

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Expression of circadian rhythm genes in gonadotropin-releasing hormone-secreting GT1-7 neurons.

Gillespie JM, Chan BP, Roy D, Cai F, Belsham DD
Endocrinology 2003 Dec;144(12):5285-92

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Circadian Transcription. Thinking outside the E-Box.

Muñoz E, Brewer M, Baler R
The Journal of biological chemistry 2002 Sep 27;277(39):36009-17

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of BMAL1 was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with BMAL1 Rabbit Polyclonal Antibody (Product # PA1-523) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of BMAL1 was done on SH-SY5Y cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with BMAL1 Rabbit Polyclonal Antibody (PA1-523, red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10, 000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Melatonin influences the phenotype of isolated P92-PSC. ( A ) Morphologic shapes of human isolated PSC cell line. Scale bars : 20 mum. ( B ) By immunofluorescence and qPCR, the MT1 and ( C and D ) clock genes (PER1, CRY1, CLOCK, and BMAL1) are decreased in isolated PSC cell lines treated with melatonin compared with vehicle-treated PSC cells. Original magnification, 20x; scale bars : 20 mum. Data are means +- SEM of 2 evaluations from 3 cumulative preparations of in vitro experiments. Each dot represents 1 value in data set. * P < .05 vs PSC.