Key features and details
- Rabbit polyclonal to Hes1
- Suitable for: WB, IHC-P, ICC/IF
- Reacts with: Human
- Isotype: IgG
Product nameAnti-Hes1 antibody
See all Hes1 primary antibodies
DescriptionRabbit polyclonal to Hes1
Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Human
Predicted to work with: Rat, Rabbit, Horse, Chicken, Guinea pig, Cow, Pig, Zebrafish
- WB: HepG2 whole cell lysate; Ls174 cells; murine embryonic cells. IHC-P: Human liver. ICC/IF: HepG2 cells; Ls174T cells.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferpH: 7.2
Preservative: 0.09% Sodium azide
Constituents: 2% Sucrose, PBS
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab49170 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1.25 µg/ml. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.|
|IHC-P||Use a concentration of 4 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ICC/IF||Use a concentration of 5 µg/ml.|
FunctionTranscriptional repressor of genes that require a bHLH protein for their transcription. May act as a negative regulator of myogenesis by inhibiting the functions of MYOD1 and ASH1. Binds DNA on N-box motifs: 5'-CACNAG-3' with high affinity and on E-box motifs: 5'-CANNTG-3' with low affinity.
Sequence similaritiesContains 1 basic helix-loop-helix (bHLH) domain.
Contains 1 Orange domain.
DomainHas a particular type of basic domain (presence of a helix-interrupting proline) that binds to the N-box (CACNAG), rather than the canonical E-box (CANNTG).
The C-terminal WRPW motif is a transcriptional repression domain necessary for the interaction with Groucho/TLE family members, transcriptional corepressors recruited to specific target DNA by Hairy-related proteins.
The bHLH, as well as cooperation between the central Orange domain and the C-terminal WRPW motif, is required for transcriptional repressor activity.
- Information by UniProt
- bHLHb39 antibody
- C-HAIRY1 antibody
- c-hairy1A antibody
Anti-Hes1 antibody (ab49170) at 1 µg/ml + HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 30 kDa
Observed band size: 30 kDa
Additional bands at: 17 kDa, 70 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 30 seconds
ab49170 (4µg/ml) staining Hes1 in human liver using an automated system (DAKO Autostainer Plus). Using this protocol there is weak cytoplasmic staining in plates of hepatic cells. However there is stronger nuclear staining in the plates of hepatic cells and hepatic arteries.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
ICC/IF image of ab49170 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab49170, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Western Blot using lysate prepared from Hes1 transfected Ls174 cells. ab49170 used at a 1µg/ml.
Western blot using lysate prepared from murine embryonic cells.
ab49170 used at a 1/800 dilution and the secondary was used at a 1/5000 dilution.
ab49170 at 50µg/ml staining Hes1 in Ls174T cells by Immunocytochemistry/ Immunofluorescence.
ab49170 has been referenced in 13 publications.
- Zhao Z et al. SPHK1 promotes metastasis of thyroid carcinoma through activation of the S1P/S1PR3/Notch signaling pathway. Exp Ther Med 15:5007-5016 (2018). PubMed: 29805524
- Wang YC et al. Notch Signaling Pathway Is Inhibited in the Development of Barrett's Esophagus: An In Vivo and In Vitro Study. Can J Gastroenterol Hepatol 2018:4149317 (2018). PubMed: 29785394
- Man J et al. Hypoxic Induction of Vasorin Regulates Notch1 Turnover to Maintain Glioma Stem-like Cells. Cell Stem Cell 22:104-118.e6 (2018). PubMed: 29198941
- Kato R et al. Dipeptide ?-secretase inhibitor treatment enhances the anti-tumor effects of cisplatin against gastric cancer by suppressing cancer stem cell properties. Oncol Lett 16:5426-5432 (2018). PubMed: 30250614
- Ziller MJ et al. Dissecting the Functional Consequences of De Novo DNA Methylation Dynamics in Human Motor Neuron Differentiation and Physiology. Cell Stem Cell 22:559-574.e9 (2018). PubMed: 29551301