Recombinant Anti-Sumo 1 antibody [EP298] (ab133352)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP298] to Sumo 1
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Sumo 1 antibody [EP298]
See all Sumo 1 primary antibodies -
Description
Rabbit monoclonal [EP298] to Sumo 1 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WBmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human Sumo 1 aa 1-100. The exact sequence is proprietary.
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Positive control
- WB- HeLa whole cell lysate (ab150035), MCF-7 whole cell lysate, A549 whole cell lysate, NIH/3T3 whole cell lysate, C6 whole cell lysate, HL-60 cell lysate and A431 cell lysate. IHC- Human papillary cell thyroid gland carcinoma tissue, Human Breast carcinoma tissue, Human Glioma tissue, Human Urinary bladder transitional carcinoma tissue and Human tonsil tissue.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.21% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP298 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab133352 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 |
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IHC-P | (1) |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB |
1/1000 - 1/10000. Predicted molecular weight: 11.5 kDa.
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Notes |
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IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
1/1000 - 1/10000. Predicted molecular weight: 11.5 kDa. |
Target
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Function
Ubiquitin-like protein that can be covalently attached to proteins as a monomer or a lysine-linked polymer. Covalent attachment via an isopeptide bond to its substrates requires prior activation by the E1 complex SAE1-SAE2 and linkage to the E2 enzyme UBE2I, and can be promoted by E3 ligases such as PIAS1-4, RANBP2 or CBX4. This post-translational modification on lysine residues of proteins plays a crucial role in a number of cellular processes such as nuclear transport, DNA replication and repair, mitosis and signal transduction. Involved for instance in targeting RANGAP1 to the nuclear pore complex protein RANBP2. Polymeric SUMO1 chains are also susceptible to polyubiquitination which functions as a signal for proteasomal degradation of modified proteins. May also regulate a network of genes involved in palate development. -
Involvement in disease
Defects in SUMO1 are the cause of non-syndromic orofacial cleft type 10 (OFC10) [MIM:613705]; also called non-syndromic cleft lip with or without cleft palate 10. OFC10 is a birth defect consisting of cleft lips with or without cleft palate. Cleft lips are associated with cleft palate in two-third of cases. A cleft lip can occur on one or both sides and range in severity from a simple notch in the upper lip to a complete opening in the lip extending into the floor of the nostril and involving the upper gum. Note=A chromosomal aberation involving SUMO1 is the cause of OFC10. Translocation t(2;8)(q33.1;q24.3). The breakpoint occurred in the SUMO1 gene and resulted in haploinsufficiency confirmed by protein assays. -
Sequence similarities
Belongs to the ubiquitin family. SUMO subfamily.
Contains 1 ubiquitin-like domain. -
Post-translational
modificationsCleavage of precursor form by SENP1 or SENP2 is necessary for function.
Polymeric SUMO1 chains undergo polyubiquitination by RNF4. -
Cellular localization
Nucleus membrane. Nucleus speckle. Cytoplasm. Recruited by BCL11A into the nuclear body. - Information by UniProt
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Database links
- Entrez Gene: 7341 Human
- Entrez Gene: 22218 Mouse
- Entrez Gene: 301442 Rat
- Omim: 601912 Human
- SwissProt: P63165 Human
- SwissProt: P63166 Mouse
- SwissProt: Q5I0H3 Rat
- Unigene: 596171 Human
see all -
Alternative names
- DAP1 antibody
- GAP modifying protein 1 antibody
- GAP-modifying protein 1 antibody
see all
Images
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Sumo 1 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: MCF-7 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab133352 observed at 16 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab133352 was shown to recognize Sumo 1 when Sumo 1 knockout samples were used, along with additional cross-reactive bands. Wild-type and Sumo 1 knockout samples were subjected to SDS-PAGE. ab133352 and ab8245 (loading control to GAPDH) were diluted 1/000 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. -
Immunohistochemical staining of paraffin-embedded human thyroid carcinoma sections labelling Sumo 1 with purified ab133352 at dilution of 1:100. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
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All lanes : Anti-Sumo 1 antibody [EP298] (ab133352) at 1/10000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma) whole cell lysate
Lane 2 : MCF-7 (human breast carcinoma) whole cell lysate
Lane 3 : A549 (human lung carcinoma) whole cell lysate
Lane 4 : NIH/3T3 (mouse embryo) whole cell lysate
Lane 5 : C6 (rat glioma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Predicted band size: 11.5 kDa
Observed band size: 18-90 kDa why is the actual band size different from the predicted?
Blocking and diluting buffer 5% NFDM/TBST -
All lanes : Anti-Sumo 1 antibody [EP298] (ab133352) at 1/1000 dilution (unpurified)
Lane 1 : HeLa cell lysate
Lane 2 : HL60 cell lysate
Lane 3 : A431 cell lysate
Lane 4 : NIH3T3 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 11.5 kDa -
Immunohistochemical analysis of paraffin-embedded Human papillary cell thyroid gland carcinoma tissue labelling Sumo 1 with un-purified ab133352 at 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin embedded Human Breast carcinoma tissue labelling Sumo 1 with un-purified ab133352 at dilution of 1/100-1/250.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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All lanes : Anti-Sumo 1 antibody [EP298] (ab133352) at 1/1000 dilution
Lane 1 :Recombinant Human Sumo 1 protein (ab140417) at 0.01 µg
Lane 2 :Recombinant Human Sumo 2 protein (ab140420) at 0.01 µg
Lane 3 : Recombinant Human Sumo 3 protein (ab140414) at 0.01 µg
Lane 4 :Recombinant Human Sumo 4 protein (ab157025) at 0.1 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Predicted band size: 11.5 kDa
Observed band size: 16 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking buffer: 5% NFDM/TBST
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Immunohistochemical analysis of paraffin embedded Human Glioma tissue labelling Sumo 1 with un-purified ab133352 at dilution of 1/100-1/250.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin embedded Human Urinary bladder transitional carcinoma tissue labelling Sumo 1 with un-purified ab133352 at dilution of 1/100-1/250.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin embedded normal Human tonsil tissue labelling Sumo 1 with un-purified ab133352 at dilution of 1/100-1/250.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (13)
ab133352 has been referenced in 13 publications.
- Xia L et al. SUMOylation disassembles the tetrameric pyruvate kinase M2 to block myeloid differentiation of leukemia cells. Cell Death Dis 12:101 (2021). PubMed: 33473116
- Lin Y et al. Hypoxia activates SUMO-1-HIF-1a signaling pathway to upregulate pro-inflammatory cytokines and permeability in human tonsil epithelial cells. Life Sci 276:119432 (2021). PubMed: 33794253
- Clemens Böckelmann L et al. Concomitant constitutive LNK and NFE2 mutation with loss of sumoylation in a case of hereditary thrombocythemia. Haematologica 106:1158-1162 (2021). PubMed: 32554556
- Xu D et al. Effects of the E1 activating enzyme UBA2 on porcine oocyte maturation, apoptosis, and embryonic development in vitro. Anim Sci J 92:e13548 (2021). PubMed: 33835647
- Lin H et al. β-Cell Knockout of SENP1 Reduces Responses to Incretins and Worsens Oral Glucose Tolerance in High-Fat Diet-Fed Mice. Diabetes 70:2626-2638 (2021). PubMed: 34462260