Anti-SUFU antibody (ab28083)
Key features and details
- Rabbit polyclonal to SUFU
- Suitable for: ICC/IF, WB
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-SUFU antibody
See all SUFU primary antibodies -
Description
Rabbit polyclonal to SUFU -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat, Chicken, Dog
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Immunogen
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Positive control
- WB: HeLa, LNCaP, HEK293T, Jurkat, A431 cell lysates; Recombinant human SUFU protein (ab113584); Mouse testis tissue lysate. ICC/IF: HeLa 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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Immunizing Peptide (Blocking)
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Isotype control
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KO cell lines
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KO cell lysates
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KO cell pellets
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Positive Controls
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab28083 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 Use a concentration of 5 µg/ml. WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa). Target
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Function
Negative regulator in the hedgehog signaling pathway. Down-regulates GLI1-mediated transactivation of target genes (PubMed:15367681, PubMed:24311597, PubMed:24217340). Down-regulates GLI2-mediated transactivation of target genes (PubMed:24311597, PubMed:24217340). Part of a corepressor complex that acts on DNA-bound GLI1. May also act by linking GLI1 to BTRC and thereby targeting GLI1 to degradation by the proteasome. Sequesters GLI1, GLI2 and GLI3 in the cytoplasm, this effect is overcome by binding of STK36 to both SUFU and a GLI protein (PubMed:10806483, PubMed:24217340). Negative regulator of beta-catenin signaling. Regulates the formation of either the repressor form (GLI3R) or the activator form (GLI3A) of the full length form of GLI3 (GLI3FL). GLI3FL is complexed with SUFU in the cytoplasm and is maintained in a neutral state. Without the Hh signal, the SUFU-GLI3 complex is recruited to cilia, leading to the efficient processing of GLI3FL into GLI3R. When Hh signaling is initiated, SUFU dissociates from GLI3FL and the latter translocates to the nucleus, where it is phosphorylated, destabilized, and converted to a transcriptional activator (GLI3A). Required for normal embryonic development. Required for the proper formation of hair follicles and the control of epidermal differentiation. -
Tissue specificity
Ubiquitous in adult tissues. Detected in osteoblasts of the perichondrium in the developing limb of 12-week old embryos. Isoform 1 is detected in fetal brain, lung, kidney and testis. Isoform 2 is detected in fetal testis, and at much lower levels in fetal brain, lung and kidney. -
Involvement in disease
Medulloblastoma -
Sequence similarities
Belongs to the SUFU family. -
Cellular localization
Cytoplasm. Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 374118 Chicken
- Entrez Gene: 51684 Human
- Entrez Gene: 24069 Mouse
- Entrez Gene: 361769 Rat
- Omim: 607035 Human
- SwissProt: Q9UMX1 Human
- SwissProt: Q9Z0P7 Mouse
- Unigene: 404089 Human
see all -
Alternative names
- OTTHUMP00000020374 antibody
- OTTHUMP00000020377 antibody
- OTTHUMP00000020379 antibody
see all
Images
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Western blot - Anti-SUFU antibody (ab28083)All lanes : Anti-SUFU antibody (ab28083) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : SUFU knockout HEK293T cell lysate
Lane 3 : LNCaP cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 54 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab28083 observed at 58 kDa. Red - loading control ab8245 observed at 36 kDa.
ab28083 Anti-SUFU antibody was shown to specifically react with SUFU in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267282 (knockout cell lysate ab257718) was used. Wild-type and SUFU knockout samples were subjected to SDS-PAGE. ab28083 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-SUFU antibody (ab28083)Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: SUFU knockout HAP1 cell lysate (20 µg)
Lane 3: LnCaP cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)
Lanes 1 to 4: Merged signal (red and green). Green - ab28083 observed at 58 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab28083 was shown to recognize SUFU when SUFU knockout samples were used, along with additional cross-reactive bands. Wild-type and SUFU knockout samples were subjected to SDS-PAGE. ab28083 and ab8245 (loading control to GAPDH) were both diluted at 1 µg/ml and 1/10 000 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 h at room temperature before imaging. -
Western blot - Anti-SUFU antibody (ab28083)All lanes : Anti-SUFU antibody (ab28083) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 :Jurkat whole cell lysate (ab7899)
Lane 3 :A-431 whole cell lysate (ab7909)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 54 kDa
Additional bands at: 37 kDa (possible cleavage fragment), 37 kDa (possible cross reactivity) -
Western blot - Anti-SUFU antibody (ab28083)Anti-SUFU antibody (ab28083) at 1 µg/ml + Testis (Mouse) Tissue Lysate - normal tissue at 10 µg
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 54 kDa -
Immunocytochemistry/ Immunofluorescence - Anti-SUFU antibody (ab28083)ICC/IF image of ab28083 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab28083, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).
Protocols
References (4)
ab28083 has been referenced in 4 publications.
- Xu F et al. LIFR-AS1 modulates Sufu to inhibit cell proliferation and migration by miR-197-3p in breast cancer. Biosci Rep 39:N/A (2019). PubMed: 31127025
- Peng Y et al. Inhibition of miR-194 suppresses the Wnt/ß-catenin signalling pathway in gastric cancer. Oncol Rep 40:3323-3334 (2018). PubMed: 30542715
- Zhou F et al. Nek2A/SuFu feedback loop regulates Gli-mediated Hedgehog signaling pathway. Int J Oncol 50:373-380 (2017). PubMed: 28035348
- Makino S et al. T396I mutation of mouse Sufu reduces the stability and activity of Gli3 repressor. PLoS One 10:e0119455 (2015). PubMed: 25760946
Images
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Western blot - Anti-SUFU antibody (ab28083)All lanes : Anti-SUFU antibody (ab28083) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : SUFU knockout HEK293T cell lysate
Lane 3 : LNCaP cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 54 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab28083 observed at 58 kDa. Red - loading control ab8245 observed at 36 kDa.
ab28083 Anti-SUFU antibody was shown to specifically react with SUFU in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267282 (knockout cell lysate ab257718) was used. Wild-type and SUFU knockout samples were subjected to SDS-PAGE. ab28083 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-SUFU antibody (ab28083)
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: SUFU knockout HAP1 cell lysate (20 µg)
Lane 3: LnCaP cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)
Lanes 1 to 4: Merged signal (red and green). Green - ab28083 observed at 58 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab28083 was shown to recognize SUFU when SUFU knockout samples were used, along with additional cross-reactive bands. Wild-type and SUFU knockout samples were subjected to SDS-PAGE. ab28083 and ab8245 (loading control to GAPDH) were both diluted at 1 µg/ml and 1/10 000 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 h at room temperature before imaging. -
Western blot - Anti-SUFU antibody (ab28083)All lanes : Anti-SUFU antibody (ab28083) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 :Jurkat whole cell lysate (ab7899)
Lane 3 :A-431 whole cell lysate (ab7909)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 54 kDa
Additional bands at: 37 kDa (possible cleavage fragment), 37 kDa (possible cross reactivity) -
Western blot - Anti-SUFU antibody (ab28083)Anti-SUFU antibody (ab28083) at 1 µg/ml + Testis (Mouse) Tissue Lysate - normal tissue at 10 µg
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 54 kDa
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Immunocytochemistry/ Immunofluorescence - Anti-SUFU antibody (ab28083)ICC/IF image of ab28083 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab28083, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).
