Anti-Smg1 antibody (ab30916)
Key features and details
- Rabbit polyclonal to Smg1
- Suitable for: ICC/IF, WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Smg1 antibody
See all Smg1 primary antibodies -
Description
Rabbit polyclonal to Smg1 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Xenopus laevis
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 3600 - C-terminus of Human Smg1.
Read Abcam's proprietary immunogen policy (Peptide available as ab31816.) -
Positive control
- HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate, Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate, A431 (Human epithelial carcinoma cell line) Whole Cell Lysate.
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
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab30916 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 ICC/IF WB Application notesICC/IF: Use at 1:200 (See Abreview Feb 20 2007).
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 340 kDa (predicted molecular weight: 340 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.Target
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Function
Serine/threonine protein kinase involved in both mRNA surveillance and genotoxic stress response pathways. Recognizes the substrate consensus sequence [ST]-Q. Plays a central role in nonsense-mediated decay (NMD) of mRNAs containing premature stop codons by phosphorylating UPF1/RENT1. Recruited by release factors to stalled ribosomes together with SMG8 and SMG9 (forming the SMG1C protein kinase complex), and UPF1 to form the transient SURF (SMG1-UPF1-eRF1-eRF3) complex. In EJC-dependent NMD, the SURF complex associates with the exon junction complex (EJC) through UPF2 and allows the formation of an UPF1-UPF2-UPF3 surveillance complex which is believed to activate NMD. Also acts as a genotoxic stress-activated protein kinase that displays some functional overlap with ATM. Can phosphorylate p53/TP53 and is required for optimal p53/TP53 activation after cellular exposure to genotoxic stress. Its depletion leads to spontaneous DNA damage and increased sensitivity to ionizing radiation (IR). May activate PRKCI but not PRKCZ. -
Tissue specificity
Widely expressed, with highest level in heart and skeletal muscle. Expressed in placenta, brain, lung and spleen, but not in liver. -
Sequence similarities
Belongs to the PI3/PI4-kinase family.
Contains 1 FAT domain.
Contains 1 FATC domain.
Contains 1 HEAT repeat.
Contains 1 PI3K/PI4K domain. -
Post-translational
modificationsAutophosphorylated. -
Cellular localization
Nucleus. Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 23049 Human
- Entrez Gene: 233789 Mouse
- Omim: 607032 Human
- SwissProt: Q96Q15 Human
- SwissProt: Q8BKX6 Mouse
- Unigene: 460179 Human
- Unigene: 309053 Mouse
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Alternative names
- 61E3.4 antibody
- ATX antibody
- hSMG-1 antibody
see all
Images
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Western blot - Anti-Smg1 antibody (ab30916)All lanes : Anti-Smg1 antibody (ab30916) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Lane 2 :Jurkat whole cell lysate (ab7899) at 10 µg
Lane 3 :A-431 whole cell lysate (ab7909) at 20 µg
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution
Performed under reducing conditions.
Predicted band size: 340 kDa
Observed band size: 340 kDaThe band at 340kDa is likely to represent isoform 3 of Smg1. The immunogen is present in all three isoforms of Smg1.
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Immunocytochemistry/ Immunofluorescence - Anti-Smg1 antibody (ab30916)This image is courtesy of an Abreview submitted by Dr Kirk McManusab30916 (1/200) staining Smg1 by ICC/IF in assynchronous HeLa Cells (green). Cells were countersatined with DAPI (red) in order to show the nucleus. Secondary antibody: Goat anti-Rabbit conjugated to Cy3®. Please refer to abreview for further details. -
Immunocytochemistry/ Immunofluorescence - Anti-Smg1 antibody (ab30916)ICC/IF image of ab30916 stained HepG2 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 (ab30916, 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. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml
Protocols
Datasheets and documents
References (4)
ab30916 has been referenced in 4 publications.
- Durand S et al. Hyperphosphorylation amplifies UPF1 activity to resolve stalls in nonsense-mediated mRNA decay. Nat Commun 7:12434 (2016). WB . PubMed: 27511142
- Melero R et al. The RNA helicase DHX34 functions as a scaffold for SMG1-mediated UPF1 phosphorylation. Nat Commun 7:10585 (2016). IP . PubMed: 26841701
- Meikar O et al. An atlas of chromatoid body components. RNA 20:483-95 (2014). ICC/IF ; Mouse . PubMed: 24554440
- Hogg JR & Goff SP Upf1 senses 3'UTR length to potentiate mRNA decay. Cell 143:379-89 (2010). WB ; Human . PubMed: 21029861
Images
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Western blot - Anti-Smg1 antibody (ab30916)All lanes : Anti-Smg1 antibody (ab30916) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Lane 2 :Jurkat whole cell lysate (ab7899) at 10 µg
Lane 3 :A-431 whole cell lysate (ab7909) at 20 µg
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution
Performed under reducing conditions.
Predicted band size: 340 kDa
Observed band size: 340 kDaThe band at 340kDa is likely to represent isoform 3 of Smg1. The immunogen is present in all three isoforms of Smg1.
-
Immunocytochemistry/ Immunofluorescence - Anti-Smg1 antibody (ab30916) This image is courtesy of an Abreview submitted by Dr Kirk McManusab30916 (1/200) staining Smg1 by ICC/IF in assynchronous HeLa Cells (green). Cells were countersatined with DAPI (red) in order to show the nucleus. Secondary antibody: Goat anti-Rabbit conjugated to Cy3®. Please refer to abreview for further details.
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Immunocytochemistry/ Immunofluorescence - Anti-Smg1 antibody (ab30916)ICC/IF image of ab30916 stained HepG2 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 (ab30916, 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. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml
