Recombinant Anti-ISG15 antibody [EPR3446] - BSA and Azide free (ab248489)
![Western blot - Anti-ISG15 antibody [EPR3446] - BSA and Azide free (ab248489)](https://www.abcam.com/ps/products/248/ab248489/Images/ab248489-410675-anti-isg15-antibody-epr3446-bsa-and-azide-free-western-blot.jpg "Western blot - Anti-ISG15 antibody [EPR3446] - BSA and Azide free (ab248489)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3446] to ISG15 - BSA and Azide free
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-ISG15 antibody [EPR3446] - BSA and Azide free
See all ISG15 primary antibodies -
Description
Rabbit monoclonal [EPR3446] to ISG15 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IFmore details
Unsuitable for: Flow Cyt,IHC-P or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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General notes
ab248489 is the carrier-free version of ab133346.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
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.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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 +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading... -
Purity
Affinity purified -
Clonality
Monoclonal -
Clone number
EPR3446 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab248489 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 |
Use at an assay dependent concentration. Detects a band of approximately 15 kDa (predicted molecular weight: 18 kDa).
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| ICC/IF |
Use at an assay dependent concentration.
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| Notes |
|---|
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WB
Use at an assay dependent concentration. Detects a band of approximately 15 kDa (predicted molecular weight: 18 kDa). |
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ICC/IF
Use at an assay dependent concentration. |
Target
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Function
Ubiquitin-like protein that is conjugated to intracellular target proteins after IFN-alpha or IFN-beta stimulation. Its enzymatic pathway is partially distinct from that of ubiquitin, differing in substrate specificity and interaction with ligating enzymes. ISG15 conjugation pathway uses a dedicated E1 enzyme, but seems to converge with the Ub conjugation pathway at the level of a specific E2 enzyme. Targets include STAT1, SERPINA3G/SPI2A, JAK1, MAPK3/ERK1, PLCG1, EIF2AK2/PKR, MX1/MxA, and RIG-1. Deconjugated by USP18/UBP43. Shows specific chemotactic activity towards neutrophils and activates them to induce release of eosinophil chemotactic factors. May serve as a trans-acting binding factor directing the association of ligated target proteins to intermediate filaments. May also be involved in autocrine, paracrine and endocrine mechanisms, as in cell-to-cell signaling, possibly partly by inducing IFN-gamma secretion by monocytes and macrophages. Seems to display antiviral activity during viral infections.
In response to IFN-tau secreted by the conceptus, may ligate to and regulate proteins involved in the release of prostaglandin F2-alpha (PGF), and thus prevent lysis of the corpus luteum and maintain the pregnancy. -
Tissue specificity
Detected in lymphoid cells, striated and smooth muscle, several epithelia and neurons. -
Sequence similarities
Contains 2 ubiquitin-like domains. -
Cellular localization
Cytoplasm. Secreted. UCRP conjugates seem to be noncovalently associated with the intermediate filaments and distributed in a punctate pattern. Also secreted. - Information by UniProt
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Database links
- Entrez Gene: 9636 Human
- Omim: 147571 Human
- SwissProt: P05161 Human
- Unigene: 458485 Human
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Alternative names
- G1P2 antibody
- hUCRP antibody
- IFI 15 antibody
see all
Images
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Western blot - Anti-ISG15 antibody [EPR3446] - BSA and Azide free (ab248489)
This data was developed using ab133346, the same antibody clone in a different buffer formulation.
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: ISG15 IFN treated wild type HAP1 whole cell lysate (20 μg)
Lane 3: ISG15 (KO) whole cell lysate (20 µg)
Lane 4: ISG15 (KO) IFN alpha treated whole cell lysate (20 µg)
Lane 5: Daudi whole cell lysate (20 µg)
Lanes 1 - 5: Merged signal (red and green). Green - ab133346 observed at 18 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab133346 was shown to specifically react with ISG15 in IFN alpha1 induced wild-type HAP1cells. Loss of signal was observed when knockout cell line was used. Wild-type and HAP1 IFN treated knockout samples were subjected to SDS-PAGE. ab133346 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1000 dilution and 1/30000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-ISG15 antibody [EPR3446] - BSA and Azide free (ab248489)](https://www.abcam.com/ps/products/248/ab248489/Images/ab248489-407894-anti-isg15-antibody-epr3446-bsa-and-azide-free-western-blot.jpg)
Western blot - Anti-ISG15 antibody [EPR3446] - BSA and Azide free (ab248489)All lanes : Anti-ISG15 antibody [EPR3446] (ab133346) at 1/2000 dilution
Lane 1 : Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 10 ng/ml IFN- a1 for 16 hours whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 18 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesThis data was developed using ab133346, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
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![Immunocytochemistry/ Immunofluorescence - Anti-ISG15 antibody [EPR3446] - BSA and Azide free (ab248489)](https://www.abcam.com/ps/products/248/ab248489/Images/ab248489-1-anti-isg15-antibody-epr3446-bsa-and-azide-free-immunocytochemistry.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-ISG15 antibody [EPR3446] - BSA and Azide free (ab248489) -
![Anti-ISG15 antibody [EPR3446] - BSA and Azide free (ab248489)](https://www.abcam.com/ps/products/248/ab248489/Images/ab248489-4-benefits-of-recombinant-antibodies.png)
Anti-ISG15 antibody [EPR3446] - BSA and Azide free (ab248489)
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
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Datasheet download
Certificate of Compliance
References (0)
ab248489 has not yet been referenced specifically in any publications.
