Recombinant Anti-HIP2/LIG antibody [EP1145Y] (ab52930)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)](https://www.abcam.com/ps/products/52/ab52930/Images/ab52930-65434-anti-hip2-antibody-ep1145y-immunohistochemistry.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1145Y] to HIP2/LIG
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-HIP2/LIG antibody [EP1145Y]
See all HIP2/LIG primary antibodies -
Description
Rabbit monoclonal [EP1145Y] to HIP2/LIG -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat
-
Immunogen
Synthetic peptide within Human HIP2/LIG aa 150-250 (internal sequence). The exact sequence is proprietary.
-
Positive control
- WB: HCT116, HeLa, Jurkat and Daudi cell lysates. IHC-P: Human liver tissue.
-
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1145Y -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Positive Controls
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab52930 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 |
|---|---|---|
| WB |
1/100000. Detects a band of approximately 24 kDa (predicted molecular weight: 22 kDa).
|
|
| IHC-P | (1) |
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
| Notes |
|---|
|
WB
1/100000. Detects a band of approximately 24 kDa (predicted molecular weight: 22 kDa). |
|
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
-
Function
Accepts ubiquitin from the E1 complex and catalyzes its covalent attachment to other proteins. In vitro, in the presence or in the absence of BRCA1-BARD1 E3 ubiquitin-protein ligase complex, catalyzes the synthesis of 'Lys-48'-linked polyubiquitin chains. Does not transfer ubiquitin directly to but elongates monoubiquitinated substrate protein. Mediates the selective degradation of short-lived and abnormal proteins, such as the endoplasmic reticulum-associated degradation (ERAD) of misfolded lumenal proteins. Ubiquitinates huntingtin. May mediate foam cell formation by the suppression of apoptosis of lipid-bearing macrophages through ubiquitination and subsequence degradation of p53/TP53. Proposed to be involved in ubiquitination and proteolytic processing of NF-kappa-B; in vitro supports ubiquitination of NFKB1. In case of infection by cytomegaloviruses may be involved in the US11-dependent degradation of MHC class I heavy chains following their export from the ER to the cytosol. In case of viral infections may be involved in the HPV E7 protein-dependent degradation of RB1. -
Tissue specificity
Expressed in all tissues tested, including spleen, thymus, prostate, testis, ovary, small intestine, colon, peripheral blood leukocytes, T-lymphocytes, monocytes, granulocytes and bone marrow mononuclear cells. Highly expressed in brain, with highest levels found in cortex and striatum and at lower levels in cerebellum and brainstem. -
Pathway
Protein modification; protein ubiquitination. -
Sequence similarities
Belongs to the ubiquitin-conjugating enzyme family.
Contains 1 UBA domain. -
Post-translational
modificationsSumoylation at Lys-14 impairs catalytic activity. -
Cellular localization
Cytoplasm. - Information by UniProt
-
Database links
- Entrez Gene: 3093 Human
- Entrez Gene: 53323 Mouse
- Entrez Gene: 289623 Rat
- Omim: 602846 Human
- SwissProt: P61086 Human
- SwissProt: P61087 Mouse
- Unigene: 50308 Human
- Unigene: 725263 Human
see all -
Alternative names
- E2 25K antibody
- E2(25K) antibody
- HIP-2 antibody
see all
Images
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)
Ab52930 (1:250) staining human HIP2/LIG in human liver tissue by immunohistochemistry using paraffin embedded tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
![Western blot - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)](https://www.abcam.com/ps/products/52/ab52930/Images/ab52930-410125-ab257779-anti-hip2lig-antibody-ep1145y-western-blot-human-knockout-lysate.jpg)
Western blot - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)All lanes : Anti-HIP2/LIG antibody [EP1145Y] (ab52930) at 1/1000 dilution
Lane 1 : Wild-type HCT116 cell lysate
Lane 2 : UBE2K knockout HCT116 cell lysate
Lane 3 : Daudi cell lysate
Lane 4 : Jurkat 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: 22 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab52930 observed at 25 kDa. Red - loading control ab8245 observed at 36 kDa.
ab52930 Anti-HIP2/LIG antibody [EP1145Y] was shown to specifically react with HIP2/LIG in wild-type HCT116 cells. Loss of signal was observed when knockout cell line ab266899 (knockout cell lysate ab257779) was used. Wild-type and HIP2/LIG knockout samples were subjected to SDS-PAGE. ab52930 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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.
-
![Western blot - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)](https://www.abcam.com/ps/products/52/ab52930/Images/ab52930-393547-anti-hip2lig-antibody-ep1145y-western-blot-human-knockout-lysate.jpg)
Western blot - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)All lanes : Anti-HIP2/LIG antibody [EP1145Y] (ab52930) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : UBE2K knockout HeLa cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : Daudi 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: 22 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab52930 observed at 26 kDa. Red - loading control ab8245 observed at 36 kDa.
ab52930 Anti-HIP2/LIG antibody [EP1145Y] was shown to specifically react with HIP2/LIG in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266031 (knockout cell lysate ab257778) was used. Wild-type and HIP2/LIG knockout samples were subjected to SDS-PAGE. ab52930 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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.
-
![Western blot - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)](https://www.abcam.com/ps/products/52/ab52930/Images/ab52930-65435-anti-hip2-antibody-ep1145y-western-blot.jpg)
Western blot - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)Anti-HIP2/LIG antibody [EP1145Y] (ab52930) at 1/100000 dilution + Daudi cell lysate at 10 µg
Secondary
Goat anti-Rabbit HRP labeled at 1/2000 dilution
Predicted band size: 22 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted? -
![Anti-HIP2/LIG antibody [EP1145Y] (ab52930)](https://www.abcam.com/ps/products/52/ab52930/Images/ab52930-5-benefits-of-recombinant-antibodies.png)
Anti-HIP2/LIG antibody [EP1145Y] (ab52930)
Protocols
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.
Datasheets and documents
-
SDS download
-
Datasheet download
References (3)
ab52930 has been referenced in 3 publications.
- McManus FP et al. Quantitative SUMO proteomics reveals the modulation of several PML nuclear body associated proteins and an anti-senescence function of UBC9. Sci Rep 8:7754 (2018). PubMed: 29773808
- Su J et al. Reduction of HIP2 expression causes motor function impairment and increased vulnerability to dopaminergic degeneration in Parkinson's disease models. Cell Death Dis 9:1020 (2018). PubMed: 30282965
- Benedikt A et al. The leukemogenic AF4-MLL fusion protein causes P-TEFb kinase activation and altered epigenetic signatures. Leukemia 25:135-44 (2011). WB ; Human . PubMed: 21030982