Recombinant Anti-ASK1 antibody [EP553Y] (ab45178)
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: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-ASK1 antibody [EP553Y]
See all ASK1 primary antibodies -
Description
Rabbit monoclonal [EP553Y] to ASK1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide corresponding to Human ASK1 aa 1-100 (N terminal).
(Peptide available asab224650) -
Positive control
- IHC-P: Human lung carcinoma tisuue. WB: HeLa (ab150035), A549, and RAW 264.7 whole cell lysates. Flow Cyt (intra): HeLa cells. ICC/IF: RAW 264.7 cells.
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General notes
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
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP553Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
---|---|---|
Flow Cyt (Intra) |
1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (3) |
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.
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ICC/IF |
1/100 - 1/250.
|
Notes |
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Flow Cyt (Intra)
1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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. |
Target
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Function
Component 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 specificity
Abundantly expressed in heart and pancreas. -
Sequence similarities
Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase kinase subfamily.
Contains 1 protein kinase domain. -
Post-translational
modificationsDephosphorylated and activated by PGAM5. - Information by UniProt
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Database links
- Entrez Gene: 4217 Human
- Entrez Gene: 26408 Mouse
- Omim: 602448 Human
- SwissProt: Q99683 Human
- SwissProt: O35099 Mouse
- Unigene: 186486 Human
- Unigene: 6595 Mouse
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Alternative names
- Apoptosis signal regulating kinase 1 antibody
- Apoptosis signal-regulating kinase 1 antibody
- ASK 1 antibody
see all
Images
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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.
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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.
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Intracellular 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).
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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.
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Anti-ASK1 antibody [EP553Y] (ab45178) at 1/2000 dilution (Unpurified) + HeLa cell lysate at 10 µg
Secondary
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.
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
References (33)
ab45178 has been referenced in 33 publications.
- Guo X et al. ASK1 signaling regulates phase-specific glial interactions during neuroinflammation. Proc Natl Acad Sci U S A 119:N/A (2022). PubMed: 35101972
- Nie B et al. Dexmedetomidine alleviates hyperalgesia in arthritis rats through inhibition of the p38MAPK signaling pathway. Immunopharmacol Immunotoxicol 44:586-593 (2022). PubMed: 35445635
- He X et al. The embryonic stem cell microenvironment inhibits mouse glioma cell proliferation by regulating the PI3K/AKT pathway. Transl Cancer Res 10:487-498 (2021). PubMed: 35116278
- Nakamura T et al. The mitochondrial Ca2+ uptake regulator, MICU1, is involved in cold stress-induced ferroptosis. EMBO Rep 22:e51532 (2021). PubMed: 33822458
- Zou W et al. ASK1/p38-mediated NLRP3 inflammasome signaling pathway contributes to aberrant retinal angiogenesis in diabetic retinopathy. Int J Mol Med 47:732-740 (2021). PubMed: 33416127