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 - BSA and Azide free
- Suitable for: Flow Cyt, ICC/IF, WB
- Knockout validated
- Reacts with: Human
Product nameAnti-USP28 antibody [EPR4249(2)] - BSA and Azide free
See all USP28 primary antibodies
DescriptionRabbit monoclonal [EPR4249(2)] to USP28 - BSA and Azide free
Tested applicationsSuitable for: Flow Cyt, ICC/IF, WBmore details
Unsuitable for: IHC-P or IP
Species reactivityReacts with: Human
Synthetic peptide, corresponding to residues in Human USP28.
- 293T, HeLa, HT1376, SW480 and A431 cell lysates; Human heart tissue lysate; HeLa cells.
Ab225537 is the carrier-free version of ab126604. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.
Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
ab225537 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.
Maxpar® is a trademark of Fluidigm Canada Inc.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
Storage instructionsShipped at 4°C. Store at +4°C. Do Not Freeze.
Dissociation constant (KD)KD = 1.41 x 10 -11 M Learn more about KD
Storage bufferpH: 7.20
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab225537 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|ICC/IF||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration. Detects a band of approximately 140 kDa (predicted molecular weight: 122 kDa).|
FunctionDeubiquitinase 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 similaritiesBelongs to the peptidase C19 family. USP28 subfamily.
Contains 1 UIM (ubiquitin-interacting motif) repeat.
modificationsDegradaded upon nickel ion level or hypoxia exposure.
Phosphorylated upon DNA damage at Ser-67 and Ser-714, by ATM or ATR.
Cellular localizationNucleus > nucleoplasm.
- Information by UniProt
- Deubiquitinating enzyme 28 antibody
- KIAA1515 antibody
- Ubiquitin carboxyl terminal hydrolase 28 antibody
This WB data was generated using the same anti-USP28 antibody clone [EPR4249(2)] in a different buffer formulation (cat# ab126604).
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.
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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126604).
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab225537 has been referenced in 1 publication.
- 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