REACTIVITY | SENSITIVITY | MW (kDa) | Isotype |
---|---|---|---|
H M R Mk | Endogenous | 50 | Rabbit IgG |
Western blot analysis of extracts from various cell lines using DEK (E4S5J) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). As expected, DEK is not expressed when knocked out in 293T cells (293T DEK KO).
Learn more about how we get our images.Immunohistochemical analysis of paraffin-embedded human colon carcinoma using DEK (E4S5J) Rabbit mAb.
Learn more about how we get our images.Immunohistochemical analysis of paraffin-embedded human colon carcinoma using DEK (E4S5J) Rabbit mAb (left) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Learn more about how we get our images.Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using DEK (E4S5J) Rabbit mAb.
Learn more about how we get our images.Immunohistochemical analysis of paraffin-embedded mouse prostate using DEK (E4S5J) Rabbit mAb.
Learn more about how we get our images.Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using DEK (E4S5J) Rabbit mAb.
Learn more about how we get our images.Immunohistochemical analysis of paraffin-embedded human gastric adenocarcinoma using DEK (E4S5J) Rabbit mAb.
Learn more about how we get our images.For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody datasheet or product webpage for recommended antibody dilution.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Loading of prestained molecular weight markers (#13953, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended.
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised June 2016
Protocol Id: 263
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Do not allow slides to dry at any time during this procedure.
For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; follow with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.
posted February 2010
revised March 2016
Protocol Id: 283
Application | Dilutions |
---|---|
Western Blotting | 1:1000 |
Immunohistochemistry (Paraffin) | 1:150 - 1:600 |
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
DEK (E4S5J) Rabbit mAb recognizes endogenous levels of total DEK protein. Non-specific staining has been observed by IHC in nerve cells.
Human, Mouse, Rat, Monkey
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human DEK protein.
The protein product of the DEK oncogene is a nuclear phosphoprotein that is highly conserved among higher eukaryotic organisms and preferentially expressed in actively proliferating and/or malignant cells (1,2). DEK is an abundant, non-histone chromosomal protein that establishes and maintains heterochromatin by interacting with HP1a, enhancing HP1a binding to tri-methyl histone H3 Lys9 and stabilizing local tri-methyl histone H3 Lys9 levels (3). DEK localized to euchromatin represses transcription by interacting with transcription factors such as RelA/p65 (4). The DEK protein also associates with mRNA processing factors to regulate splicing and nuclear export (5,6).
The DEK proto-oncogene functions as a negative regulator of cellular differentiation, senescence, and apoptosis. DEK is translocated and/or over-expressed in a number of different cancers, including acute myeloid leukemia, breast cancer, cervical cancer, hepatocellular carcinoma, melanoma, and small cell lung cancer (1,2). In addition to the role of DEK in cancer biology, which is mainly related to its intracellular functions, extracellular DEK is implicated in the pathogenesis of autoimmune disorders (1,2). Circulating autoantibodies to DEK have been identified in the serum of patients with autoimmune diseases, including juvenile idiopathic arthritis, sarcoidosis, and systemic lupus erythematosus. DEK is secreted by human monocyte-derived macrophages and apoptotic T-lymphocytes and can act as a chemotactic, pro-inflammatory factor (7,8). Exogenous DEK can penetrate neighboring cells, and translocate to the nucleus to carry out its endogenous nuclear functions (9). IL-8 induced secretion of DEK from macrophages serves as a chemoattractant for peripheral blood leukocytes (7).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. SignalStain is a trademark of Cell Signaling Technology, Inc. XP is a registered trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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Product # | Size | Price |
---|---|---|
29812S | 100 µl | $ 269.0 |