PA5-34706

antibody from Invitrogen Antibodies

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

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-34706

Provider

Invitrogen Antibodies

Product name

BMAL1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: BMAL1 tansfected 293T cell, mouse brain. Predicted reactivity: Mouse (99%), Rat (99%), Xenopus laevis (96%), Pig (99%), Chicken (97%), Sheep (100%), Rhesus Monkey (100%), Bovine (100%). IHC notes, Requires antigen retrieval using heat mediated 10mM Citrate buffer (pH6.0) or Tris-EDTA buffer (pH8.0) Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.65 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of BMAL1 expression in transfected 293T cell line by BMAL1 polyclonal antibody. A: Non-transfected lysate. B: BMAL1 transfected lysate. 7.5% SDS PAGE. BMAL1 Polyclonal Antibody (Product # PA5-34706) diluted at 1:500.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using BMAL1 Polyclonal Antibody (Product # PA5-34706). Sample (50 µg of whole cell lysate). Lane A: Mouse brain . 7.5% SDS PAGE. BMAL1 Polyclonal Antibody (Product # PA5-34706) diluted at 1:1,000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of BMAL1 expression in transfected 293T cell line by BMAL1 polyclonal antibody. A: Non-transfected lysate. B: BMAL1 transfected lysate. 7.5% SDS PAGE. BMAL1 Polyclonal Antibody (Product # PA5-34706) diluted at 1:500.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-BMAL1 Polyclonal Antibody (Product # PA5-34706) and a 64 kDa band corresponding to BMAL1 was observed across cell lines and tissue extract tested. Whole cell extracts (30 µg lysate) of SH-SY5Y (Lane1), HCT 116 (Lane 2), HT-29 (Lane 3), A549 (Lane 4), Daudi (Lane 5), SK-BR-3 (Lane 6) and tissue extract of Mouse Brain (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

BMAL1 Polyclonal Antibody detects BMAL1 protein at nucleus by immunofluorescent analysis.Sample: A375 cells were fixed in 4% paraformaldehyde at RT for 15 min.Green: BMAL1 protein stained by BMAL1 Polyclonal Antibody (Product # PA5-34706) diluted at 1:500.Blue: Hoechst 33342 staining.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Arntl was performed using 70% confluent log phase NIH:OVCAR-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with BMAL1 Polyclonal Antibody (Product # PA5-34706) at 1:200 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (1:2500), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained withRhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents the low expression model Hep G2. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.