Recombinant Anti-USP28 antibody [EPR4249(2)] (ab126604)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4249(2)] to USP28
- Suitable for: Flow Cyt (Intra), WB, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-USP28 antibody [EPR4249(2)]
See all USP28 primary antibodies -
Description
Rabbit monoclonal [EPR4249(2)] to USP28 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IFmore details
Unsuitable for: IHC-P or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide, corresponding to residues in Human USP28.
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Positive control
- 293T, HeLa, HT1376, SW480 and A431 cell lysates; Human heart tissue lysate; HeLa cells.
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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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 1.41 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR4249(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab126604 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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB |
1/1000 - 1/10000. Detects a band of approximately 140 kDa (predicted molecular weight: 122 kDa).
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ICC/IF |
1/50 - 1/100.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
1/1000 - 1/10000. Detects a band of approximately 140 kDa (predicted molecular weight: 122 kDa). |
ICC/IF
1/50 - 1/100. |
Target
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Function
Deubiquitinase involved in DNA damage response checkpoint and MYC proto-oncogene stability. Involved in DNA damage induced apoptosis by specifically deubiquitinating proteins of the DNA damage pathway such as CLSPN. Also involved in G2 DNA damage checkpoint, by deubiquitinating CLSPN, and preventing its degradation by the anaphase promoting complex/cyclosome (APC/C). In contrast, it does not deubiquitinate PLK1. Specifically deubiquitinates MYC in the nucleoplasm, leading to prevent MYC degradation by the proteasome: acts by specifically interacting with isoform 1 of FBXW7 (FBW7alpha) in the nucleoplasm and counteracting ubiquitination of MYC by the SCF(FBW7) complex. In contrast, it does not interact with isoform 4 of FBXW7 (FBW7gamma) in the nucleolus, allowing MYC degradation and explaining the selective MYC degradation in the nucleolus. -
Sequence similarities
Belongs to the peptidase C19 family. USP28 subfamily.
Contains 1 UIM (ubiquitin-interacting motif) repeat. -
Post-translational
modificationsDegradaded upon nickel ion level or hypoxia exposure.
Phosphorylated upon DNA damage at Ser-67 and Ser-714, by ATM or ATR. -
Cellular localization
Nucleus > nucleoplasm. - Information by UniProt
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Database links
- Entrez Gene: 57646 Human
- Omim: 610748 Human
- SwissProt: Q96RU2 Human
- Unigene: 503891 Human
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Alternative names
- Deubiquitinating enzyme 28 antibody
- KIAA1515 antibody
- Ubiquitin carboxyl terminal hydrolase 28 antibody
see all
Images
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: USP28 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab126604 observed at 128 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab126604 was shown to recognize USP28 when USP28 knockout samples were used, along with additional cross-reactive bands. Wild-type and USP28 knockout samples were subjected to SDS-PAGE. ab126604 and ab8245 (loading control to GAPDH) were diluted 1/1000 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. -
ab126604 at 1/50 dilution staining USP28 in HeLa cells by Immunofluorescence.
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Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling USP28 with unpurified ab126604 at 1/200 dilution (1ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) (ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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All lanes : Anti-USP28 antibody [EPR4249(2)] (ab126604) at 1/1000 dilution
Lane 1 : 293T cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : HT1376 cell lysate
Lane 4 : Human heart tissue lysate
Lane 5 : SW480 cell lysate
Lane 6 : A431 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-Rabbit HRP at 1/2000 dilution
Predicted band size: 122 kDa
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (8)
ab126604 has been referenced in 8 publications.
- Ni T et al. PTBP1 drives c-Myc-dependent gastric cancer progression and stemness. Br J Cancer 128:1005-1018 (2023). PubMed: 36635500
- Zhang A et al. USP33 deubiquitinates and stabilizes HIF-2alpha to promote hypoxia response in glioma stem cells. EMBO J 41:e109187 (2022). PubMed: 35191554
- Wang H et al. USP28 and USP25 are downregulated by Vismodegib in vitro and in colorectal cancer cell lines. FEBS J 288:1325-1342 (2021). PubMed: 32578360
- Chen L et al. USP28 facilitates pancreatic cancer progression through activation of Wnt/β-catenin pathway via stabilising FOXM1. Cell Death Dis 12:887 (2021). PubMed: 34584067
- Ruiz EJ et al. USP28 deletion and small-molecule inhibition destabilizes c-MYC and elicits regression of squamous cell lung carcinoma. Elife 10:N/A (2021). PubMed: 34636321
- Liu J et al. Arginine methylation-dependent LSD1 stability promotes invasion and metastasis of breast cancer. EMBO Rep 21:e48597 (2020). PubMed: 31833203
- Müller I et al. Cancer Cells Employ Nuclear Caspase-8 to Overcome the p53-Dependent G2/M Checkpoint through Cleavage of USP28. Mol Cell 77:970-984.e7 (2020). PubMed: 31982308
- Meitinger F et al. 53BP1 and USP28 mediate p53 activation and G1 arrest after centrosome loss or extended mitotic duration. J Cell Biol 214:155-66 (2016). Human . PubMed: 27432897