Anti-Lefty antibody (ab22569)
Key features and details
- Rabbit polyclonal to Lefty
- Suitable for: WB, ICC/IF
- Reacts with: Mouse, Human, Recombinant fragment
- Isotype: IgG
Overview
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Product name
Anti-Lefty antibody
See all Lefty primary antibodies -
Description
Rabbit polyclonal to Lefty -
Host species
Rabbit -
Specificity
This antibody has been tested against recombinant protein only (human LeftyA and mouse Lefty1). I is expected to cross react with Human LeftyB as there is 100% homology of the immunogen with Human LeftyB. -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB Recombinant fragment
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Immunogen
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab22569 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanWB Recombinant fragmentApplication Abreviews Notes WB (1) ICC/IF Notes Application notesICC/IF: Use at a concentration of 1 µg/ml.
IHC-P: 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB: Use at a concentration of 1 µg/ml. Detects a bands of approximately 44, 38 and 34 kDa (predicted molecular weight: 41 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.Target
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Relevance
During vertebrate embryogenesis, a left right axis is established. Secreted growth factors of the TGF beta family, including gene products derived from nodal, lefty 1 and lefty 2, play crucial roles in establishing left right asymmetries. TGF beta (Transforming growth factor beta) is a pleiotropic cytokine that regulates growth and differentiation of diverse types of cells. TGF beta actions are directed by ligand induced activation of TGF beta receptors. Complexes formed move into the nucleus, where they act as components of a transcriptional complex. Lefty, a novel member of the TGF beta superfamily, inhibits TGF beta signaling. Lefty acts to inhibit phosphorylation of Smad2 following activation of the TGF beta receptor. Lefty also inhibits events downstream from R Smad phosphorylation. Lefty provides a repressed state of TGF beta responsive genes. The Lefty family is comprised of Lefty 1 and Lefty 2 in mouse, and Lefty A and Lefty B in humans. Members of the TGF beta superfamily require processing for their activation. Cleavage is therefore an essential step for Lefty activation. Lefty is synthesized as a large inactive precursor (41 Kda) that must be endoproteolytically processed to release the bioactive polypeptide (28 kDa and 34 kDa forms). The 28kDa form induces MAPK activity. -
Cellular localization
Secreted -
Database links
- Entrez Gene: 10637 Human
- Entrez Gene: 13590 Mouse
- Omim: 603037 Human
- SwissProt: O75610 Human
- SwissProt: Q64280 Mouse
- Unigene: 656214 Human
- Unigene: 378911 Mouse
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Alternative names
- AI450052 antibody
- Ebaf antibody
- Left right determination factor 1 antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-Lefty antibody (ab22569)ICC/IF image of ab22569 stained mouse embryonic stem cells. The cells were 4% PFA 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 (ab22569, 1µg/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).
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Western blot - Anti-Lefty antibody (ab22569)All lanes : Anti-Lefty antibody (ab22569) at 1 µg/ml
Lane 1 : Mouse Lefty1 Recombinant Protein
Lane 2 : Human LeftyA Recombinant Protein
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Rabbit IgG secondary antibody (ab28446) at 1/10000 dilution
Predicted band size: 41 kDa
Observed band size: 34,38,44 kDa why is the actual band size different from the predicted?
The mouse recombinant Lefty protein consists of two bands of ~34 and 12 kDa. Of these, ab22569 recognises the 34 kDa band. The human recombinant Lefty protein consists of three bands of 44, 38 and 34 kDa. Of these, the first two are recognised by ab22569. The protein sizes differ from the expected molecular weight of Lefty due to proteolytic processing and glycosylation.
Protocols
Datasheets and documents
References (6)
ab22569 has been referenced in 6 publications.
- Harpelunde Poulsen K et al. Influence of Nodal signalling on pluripotency factor expression, tumour cell proliferation and cisplatin-sensitivity in testicular germ cell tumours. BMC Cancer 20:349 (2020). PubMed: 32326899
- Zhang P et al. Dissecting the Single-Cell Transcriptome Network Underlying Gastric Premalignant Lesions and Early Gastric Cancer. Cell Rep 27:1934-1947.e5 (2019). PubMed: 31067475
- Xu C et al. Lefty1 alleviates renal tubulointerstitial injury in mice with unilateral ureteral obstruction. Mol Med Rep 13:901-8 (2016). IHC-P ; Mouse . PubMed: 26647776
- Yoon SJ et al. HEB associates with PRC2 and SMAD2/3 to regulate developmental fates. Nat Commun 6:6546 (2015). WB, IP ; Mouse . PubMed: 25775035
- Sun G et al. Lefty inhibits glioma growth by suppressing Nodal-activated Smad and ERK1/2 pathways. J Neurol Sci 347:137-42 (2014). PubMed: 25304053
- Barroso-delJesus A et al. The Nodal inhibitor Lefty is negatively modulated by the microRNA miR-302 in human embryonic stem cells. FASEB J 25:1497-508 (2011). WB, IHC (PFA fixed) ; Human . PubMed: 21266536
Images
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Immunocytochemistry/ Immunofluorescence - Anti-Lefty antibody (ab22569)ICC/IF image of ab22569 stained mouse embryonic stem cells. The cells were 4% PFA 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 (ab22569, 1µg/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).
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Western blot - Anti-Lefty antibody (ab22569)All lanes : Anti-Lefty antibody (ab22569) at 1 µg/ml
Lane 1 : Mouse Lefty1 Recombinant Protein
Lane 2 : Human LeftyA Recombinant Protein
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Rabbit IgG secondary antibody (ab28446) at 1/10000 dilution
Predicted band size: 41 kDa
Observed band size: 34,38,44 kDa why is the actual band size different from the predicted?
The mouse recombinant Lefty protein consists of two bands of ~34 and 12 kDa. Of these, ab22569 recognises the 34 kDa band. The human recombinant Lefty protein consists of three bands of 44, 38 and 34 kDa. Of these, the first two are recognised by ab22569. The protein sizes differ from the expected molecular weight of Lefty due to proteolytic processing and glycosylation.
