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Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse, Rat

Applications

Chromatin Immunoprecipitation, Chromatin Immunoprecipitation (ChIP), Chromatin Immunoprecipitation Sequencing, ELISA, Flow Cytometry, Immunoblotting, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1 mg/ml

Product Summary for DEC1 Antibody - BSA Free

Immunogen

Synthetic peptide made to a C-terminal region of human DEC1 (between amino acids 350-412) [UniProt O14503]

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 26834156).

Localization

Nucleus

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

46 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.

Scientific Data Images for DEC1 Antibody - BSA Free

Western Blot: DEC1 Antibody - BSA Free [NB100-1800] - Western Blot Image of anti-DEC1. Whole cell protein from A431 (lane 1) and PC3 (lane 2) was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2 ug/ml anti-DEC1 in 1% milk, and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.
Immunocytochemistry/Immunofluorescence: DEC1 Antibody - BSA Free [NB100-1800] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-DEC1 (NB100-1800) at a 1:200 dilution overnight at 4C and detected with and anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin was used as a co-stain at a 1:1000 dilution and detected with and anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Immunohistochemistry: DEC1 Antibody - BSA Free [NB100-1800] - Immunohistochemical detection of DEC1 in myocardial and stromal cells. Representative images of one WT heart treated with TAC and sham at four weeks. The black square shows representative large images, magnification 400x. Image collected and cropped by CiteAb from the following publication (https://www.mdpi.com/1422-0067/20/19/4967) licensed under a CC-BY license.

Applications for DEC1 Antibody - BSA Free

Application
Recommended Usage

Chromatin Immunoprecipitation

reported in scientific literature (PMID 29715265)

Chromatin Immunoprecipitation (ChIP)

reported in scientific literature (PMID 31061528)

Chromatin Immunoprecipitation Sequencing

reported in scientific literature (PMID 31061528)

ELISA

reported in scientific literature (PMID 31061528)

Flow Cytometry

2-5 ug/ml

Immunoblotting

1:500-1:1500

Immunocytochemistry/ Immunofluorescence

1:500 - 1:1000

Immunohistochemistry

1:200 - 1:500

Immunohistochemistry-Paraffin

1:200 - 1:500

Simple Western

1:50

Western Blot

1:500-1:1500
Application Notes
In Western Blot, a band is seen approx. 49 kDa. In ICC/IF punctate nuclear staining is observed.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 5 using NB100-1800 in the following applications:

Published Applications

Read 41 publications using NB100-1800 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: DEC1

The basic helix-loop-helix (bHLH) transcription factor DEC1 (differentially expressed in chondrocytes protein 1) is wildly expressed in most normal tissues in a proportion of cells, whereas in tumors, its expression gets upregulated (potentially by hypoxia) in several malignancies such as breast, stomach, colon, lung, liver cancer and pancreatic ductal adenocarcinoma. The gene expression of DEC1 is regulated by various extracellular stimuli in a cell type-specific manner. Localized in the nucleus, it can homodimerize or heterodimerize with TCF3/E47, and interacts with ubiquitin-conjugating enzyme UBE2I/UBC9. DEC1 has emerged as one of the major circadian molecules alongwith its involvement in homeostasis of metabolism and energy. DEC1 has been shown to play a key role in various biological phenomena such as circadian rhythms, neurogenesis, neuroregulation, chondrogenesis, cellular growth, carcinogenesis, immune response balance as well as cellular senescence.

Alternate Names

basic helix-loop-helix domain containing, class B, 2, basic helix-loop-helix family, member e40, bHLHb2, bHLHe40, Class B basic helix-loop-helix protein 2, class E basic helix-loop-helix protein 40, DEC1HLHB2, differentially expressed in chondrocytes 1, Differentially expressed in chondrocytes protein 1, differentiated embryo chondrocyte expressed gene 1, Enhancer-of-split and hairy-related protein 2, SHARP2, SHARP-2, Stimulated by retinoic acid gene 13 protein, STRA13FLJ99214, Stra14

Entrez Gene IDs

8553 (Human)

Gene Symbol

BHLHE40

UniProt

Product Documents for DEC1 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for DEC1 Antibody - BSA Free

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.

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Protocols

View specific protocols for DEC1 Antibody - BSA Free (NB100-1800):

[[URL:https://www.novusbio.com/products/dec1-antibody_nb100-1800]][[Caption:D… Antibody]]
Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
[[URL:https://www.novusbio.com/products/dec1-antibody_nb100-1800]][[Caption:D… Antibody]]
Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 25 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute anti-DEC1 primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

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