Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP553Y] to ASK1
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Human
Product nameAnti-ASK1 antibody [EP553Y]
See all ASK1 primary antibodies
DescriptionRabbit monoclonal [EP553Y] to ASK1
Tested applicationsSuitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
Species reactivityReacts with: Mouse, Human
Synthetic peptide corresponding to Human ASK1 aa 1-100 (N terminal).
(Peptide available as
- IHC-P: Human lung carcinoma tisuue. WB: HeLa (ab150035), A549, and RAW 264.7 whole cell lysates. Flow Cyt: HeLa cells. ICC/IF: RAW 264.7 cells.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.5% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab45178 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000. Detects a band of approximately 155 kDa (predicted molecular weight: 155 kDa).
For unpurified use at 1/2000
|IHC-P||1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ICC/IF||1/100 - 1/250.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionComponent of a protein kinase signal transduction cascade. Phosphorylates and activates MAP2K4 and MAP2K6, which in turn activate the JNK and p38 MAP kinases, respectively. Overexpression induces apoptotic cell death.
Tissue specificityAbundantly expressed in heart and pancreas.
Sequence similaritiesBelongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase kinase subfamily.
Contains 1 protein kinase domain.
modificationsDephosphorylated and activated by PGAM5.
- Information by UniProt
- Apoptosis signal regulating kinase 1 antibody
- Apoptosis signal-regulating kinase 1 antibody
- ASK 1 antibody
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: ASK1 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: A549 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab45178 observed at 155, 160 kDa. Red - loading control, ab1240, observed at 124 kDa.
Unpurified ab45178 was shown to recognize ASK1 when ASK1 knockout samples were used, along with additional cross-reactive bands. Wild-type and ASK1 knockout samples were subjected to SDS-PAGE. ab45178 and ab1240 (loading control to Vinculin) were diluted to 1/2000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence analysis of RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells labeling ASK1 with Purified ab45178 at 1:100 (9.9 µg/ml). Cells were fixed in 100% Methanol. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling ASK1 with purified ab45178 at 1:100 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Unpurified ab45178 (1/10) staining human ASK1 in human lung carcinoma by immunohistochemistry using paraffin embedded tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Anti-ASK1 antibody [EP553Y] (ab45178) at 1/2000 dilution (Unpurified) + HeLa cell lysate at 10 µg
Goat anti-rabbit, HRP labelled at 1/2000 dilution
Predicted band size: 155 kDa
Observed band size: 155 kDa
Overlay histogram showing HeLa cells stained with unpurified ab45178 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab45178, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit DyLight® 488 (IgG H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab45178 has been referenced in 22 publications.
- Challa TD et al. Liver ASK1 protects from non-alcoholic fatty liver disease and fibrosis. EMBO Mol Med N/A:N/A (2019). PubMed: 31171651
- Giuliano CJ et al. MELK expression correlates with tumor mitotic activity but is not required for cancer growth. Elife 7:N/A (2018). PubMed: 29417930
- Cheng R et al. ß-TrCP-dependent degradation of ASK1 suppresses the induction of the apoptotic response by oxidative stress. Biochim Biophys Acta Gen Subj 1862:2271-2280 (2018). PubMed: 30031111
- Stöcker S et al. A role for 2-Cys peroxiredoxins in facilitating cytosolic protein thiol oxidation. Nat Chem Biol 14:148-155 (2018). PubMed: 29251718
- Leestemaker Y et al. Proteasome Activation by Small Molecules. Cell Chem Biol 24:725-736.e7 (2017). PubMed: 28552582