Product datasheet
Anti-ISG15 antibody ab36765
3 Images
Overview
Product name
Anti-ISG15 antibody
Description
Rabbit polyclonal to ISG15
Tested applications
WB, ICC/IF, IHC-P
Species reactivity
Reacts with: Human
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human
ISG15.Read Abcam's proprietary immunogen policy(Peptide available as ab39552.)
Positive control
This antibody gave a positive signal in the following Whole Cell Lysates: U937, MCF7, MDA MB
361. This antibody gave a positive result in IHC in the following FFPE tissue: Human breast
fibroadenoma. This antibody gave a positive result when used in the following formaldehyde
fixed cell lines: T47D
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or 80°C. Avoid freeze / thaw cycle.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab36765 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
WB
Abreviews
Notes
Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa
(predicted molecular weight: 15 kDa).
ICC/IF
Use a concentration of 5 µg/ml.
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Application
IHC-P
Abreviews
Notes
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with
citrate buffer pH 6 before commencing with IHC staining protocol.
Target
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.
Anti-ISG15 antibody images
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ICC/IF image of ab36765 stained T47D cells.
The cells were 4% formaldehyde fixed (10
min) and then incubated in 1%BSA / 10%
normal goat serum / 0.3M glycine in 0.1%
PBS-Tween for 1h to permeabilise the cells
and block non-specific protein-protein
interactions. The cells were then incubated
with the antibody ab36765 at 5µg/ml
overnight at +4°C. The secondary antibody
(pseudo-colored green) was Alexa Fluor®
488 goat anti- rabbit (ab150081) IgG (H+L)
Immunocytochemistry/ Immunofluorescence -
preadsorbed, used at a 1/1000 dilution for 1h.
Anti-ISG15 antibody (ab36765)
Alexa Fluor® 594 WGA was used to label
plasma membranes (pseudo-colored red) at
a 1/200 dilution for 1h at room temperature.
DAPI was used to stain the cell nuclei
(pseudo-colored blue) at a concentration of
1.43µM for 1hour at room temperature.
IHC image of ISG15 staining in Human breast
fibroadenoma formalin fixed paraffin
embedded tissue section, performed on a
Leica Bondâ„¢ system using the standard
protocol F. The section was pre-treated using
heat mediated antigen retrieval with sodium
citrate buffer (pH6, epitope retrieval solution
1) for 20 mins. The section was then
incubated with ab36765, 1µg/ml, for 15 mins
at room temperature and detected using an
Immunohistochemistry (Formalin/PFA-fixed
paraffin-embedded sections) - Anti-ISG15 antibody
(ab36765)
HRP conjugated compact polymer system.
DAB was used as the chromogen. The
section was then counterstained with
haematoxylin and mounted with DPX.
For other IHC staining systems (automated
and non-automated) customers should
optimize variable parameters such as antigen
retrieval conditions, primary antibody
concentration and antibody incubation times.
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All lanes : Anti-ISG15 antibody (ab36765) at
1 µg/ml
Lane 1 : U937 (Human leukemic monocyte
lymphoma cell line) Whole Cell Lysate
(ab3959)
Lane 2 : U937 (Human leukemic monocyte
lymphoma cell line) Nuclear Lysate - IFN
alpha treated (ab14907)
Lane 3 : MCF7 (Human breast
Western blot - ISG15 antibody (ab36765)
adenocarcinoma cell line) Whole Cell Lysate
Lane 4 : MDA MB 361 (Human breast
adenocarcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - PreAdsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 15 kDa
Observed band size : 17 kDa
We are unsure as to the identity of the
additional bands.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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