Recombinant Anti-UFC1 antibody [EPR15014] (ab189251)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR15014] to UFC1
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-UFC1 antibody [EPR15014]
See all UFC1 primary antibodies -
Description
Rabbit monoclonal [EPR15014] to UFC1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human fetal liver tissue lysate and HEK-293T, HAP1, A549, U-87 MG, MCF7, C6, PC12, Raw 264.7 and NIH 3T3 cell lysates. IHC-P: Human colon and hepatocellular carcinoma tissues.
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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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR15014 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab189251 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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WB |
1/10000 - 1/50000. Detects a band of approximately 19 kDa (predicted molecular weight: 19 kDa).
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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WB
1/10000 - 1/50000. Detects a band of approximately 19 kDa (predicted molecular weight: 19 kDa). |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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Function
E2-like enzyme which forms an intermediate with UFM1 via a thioester linkage. -
Sequence similarities
Belongs to the ubiquitin-conjugating enzyme family. UFC1 subfamily. - Information by UniProt
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Database links
- Entrez Gene: 51506 Human
- Entrez Gene: 66155 Mouse
- Entrez Gene: 445268 Rat
- Omim: 610554 Human
- SwissProt: Q9Y3C8 Human
- SwissProt: Q9CR09 Mouse
- SwissProt: Q6BBI8 Rat
- Unigene: 301412 Human
see all -
Alternative names
- CGI-126 antibody
- HSPC155 antibody
- Ubiquitin fold modifier conjugating enzyme 1 antibody
see all
Images
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All lanes : Anti-UFC1 antibody [EPR15014] (ab189251) at 1/10000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : UFC1 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 19 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab189251 observed at 20 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab189251 was shown to react with UFC1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266814 (knockout cell lysate ab257781) was used. Wild-type HEK-293T and UFC1 HEK-293T KO cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab189251 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 dilution and a 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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Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling UFC1 with ab189251 at 1/100 dilution, followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-UFC1 antibody [EPR15014] (ab189251) at 1/10000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : UFC1 knockout HAP1 cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 19 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab189251 observed at 20 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab189251 was shown to react with UFC1 in wild-type HAP1 cells in Western blot. Loss of signal was observed when UFC1 knockout sample was used. Wild-type HAP1 and UFC1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab189251 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-UFC1 antibody [EPR15014] (ab189251) at 1/20000 dilution
Lane 1 : Human fetal liver lysate
Lane 2 : A549 cell lysate
Lane 3 : U-87 MG cell lysate
Lane 4 : MCF7 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-rabbit IgG, (H+L), peroxidase conjugate at 1000 µg
Predicted band size: 19 kDa
Observed band size: 19 kDa -
All lanes : Anti-UFC1 antibody [EPR15014] (ab189251) at 1/5000 dilution
Lane 1 : C6 cell lysate
Lane 2 : Raw 264.7 cell lysate
Lane 3 : PC12 cell lysate
Lane 4 : NIH 3T3 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit IgG, (H+L), peroxidase conjugate at 1/1000 dilution
Predicted band size: 19 kDa
Observed band size: 19 kDa -
Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling UFC1 with ab189251 at 1/100 dilution, followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (3)
ab189251 has been referenced in 3 publications.
- Wesch N et al. A Concerted Action of UBA5 C-Terminal Unstructured Regions Is Important for Transfer of Activated UFM1 to UFC1. Int J Mol Sci 22:N/A (2021). PubMed: 34299007
- Nahorski MS et al. Biallelic UFM1 and UFC1 mutations expand the essential role of ufmylation in brain development. Brain 141:1934-1945 (2018). PubMed: 29868776
- Colin E et al. Biallelic Variants in UBA5 Reveal that Disruption of the UFM1 Cascade Can Result in Early-Onset Encephalopathy. Am J Hum Genet 99:695-703 (2016). PubMed: 27545681