Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4888] to ULK3
- Suitable for: WB, Flow Cyt, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Human
Product nameAnti-ULK3 antibody [EPR4888]
See all ULK3 primary antibodies
DescriptionRabbit monoclonal [EPR4888] to ULK3
Tested applicationsSuitable for: WB, Flow Cyt, ICC/IF, IHC-Pmore details
Unsuitable for: IP
Species reactivityReacts with: Human
Does not react with: Mouse, Rat
Synthetic peptide within Human ULK3 aa 400-500 (C terminal). The exact sequence is proprietary.
Database link: Q6PHR2-3
- IHC-P: Human bladder carcinoma and human colon tissue; WB: Wild-type HAP1 whole cell lysate; HEK-293 and LNCaP whole cell lysates. Human heart and testis tissue lysates. ICC/IF: LNCaP cells; Flow Cyt: LNCaP cells.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol, 59% PBS
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab124947 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 - 1/10000. Predicted molecular weight: 53 kDa.|
|Flow Cyt||1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|IHC-P||1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
FunctionSerine/threonine protein kinase which enhances GLI1 and GLI2 transcriptional activity and consequently positively regulates GLI-dependent SHH signaling. May exert this function by promoting GLI1 nuclear localization. Phosphorylates in vitro GLI2, as well as GLI1 and GLI3, although less efficiently.
Tissue specificityWidely expressed. Highest levels observed in fetal brain. In adult tissues, high levels in brain, liver and kidney, moderate levels in testis and adrenal gland and low levels in heart, lung, stomach, thymus, prostate and placenta. In the brain, highest expression in the hippocampus, high levels also detected in the cerebellum, olfactory bulb and optice nerve. In the central nervous system, lowest levels in the spinal cord.
Sequence similaritiesBelongs to the protein kinase superfamily. Ser/Thr protein kinase family. APG1/unc-51/ULK1 subfamily.
Contains 2 MIT domains.
Contains 1 protein kinase domain.
modificationsAutophosphorylated in vitro.
- Information by UniProt
FormThere are 3 isoforms produced by alternative splicing.
- DKFZP434C131 antibody
- FLJ90566 antibody
- Serine/threonine-protein kinase ULK3 antibody
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: ULK3 knockout HAP1 whole cell lysate (20 µg)
Lane 3: Hek293 whole cell lysate (20 µg)
Lanes 1 - 3: Merged signal (red and green). Green - ab124947 observed at 55 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab124947 (unpurified) was shown to specifically react with ULK3 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when ULK3 knockout samples were examined. Wild-type and ULK3 knockout samples were subjected to SDS-PAGE. Ab124947 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labeling ULK3 with purified ab124947 at 1/50 dilution (7.84 µg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Immunocytochemistry/Immunofluorescence analysis of LNCaP (Human prostate carcinoma epithelial cell) cells labeling ULK3 with Purified ab124947 at 1:50 dilution (7.8 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. 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 (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
All lanes : Anti-ULK3 antibody [EPR4888] (ab124947) at 1/1000 dilution (Purified)
Lane 1 : Human heart lysate
Lane 2 : Human testis lysate
Lane 3 : LNCaP (Human prostate carcinoma epithelial cell) whole cell lysate
Lane 4 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
Blocking/Diluting buffer: 5% NFDM/TBST
Flow Cytometry analysis of LNCaP (Human prostate carcinoma epithelial cell) cells, labeling ULK3 with Purified ab124947 at 1:40 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
ab124947 (unpurified), at a 1/50 dilution, staining ULK3 in paraffin embedded Human bladder carcinoma tissue by Immunohistochemistry. Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
ab124947 has been referenced in 1 publication.
- Hutterer C et al. Profiling of the kinome of cytomegalovirus-infected cells reveals the functional importance of host kinases Aurora A, ABL and AMPK. Antiviral Res 99:139-48 (2013). WB ; Human . PubMed: 23648710