Western blot analysis of extracts from various cell lines using KLF4 antibody.
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Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 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.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
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: 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 October 2016
Protocol Id: 10
KLF4 Antibody detects endogenous levels of total KLF4 protein.Species Reactivity:
Human, Mouse, Monkey
Polyclonal antibodies are produced by immunizing animals with synthetic peptide corresponding to amino acid sequence at the amino terminus of human KLF4. Antibodies are purified by Protein A and peptide affinity chromatography.
KLF4 is a member of the erythroid Kruppel-like factor (EKLF) multigene family that is highly expressed in the differentiating layers of the epidermis (1, 2). KLF4 plays a critical role in the differentiation of epithelial cells and is essential for normal gastric homeostasis (2,3). Depending on the target gene, KLF4 can function as both a repressor and activator of transcription (4). Research studies suggest this protein may function as either a tumor suppressor or an oncogene depending on tumor type, with up-regulation in human squamous cell carcinoma of the head and neck and down-regulation in colorectal carcinoma (5,6). The in vitro reprogramming of somatic cells to an embryonic-like state has been achieved by retroviral transduction of four factors: Oct-3/4, Sox2, c-Myc, and KLF4 (7). These induced pluripotent stem cells (iPS) are of great therapeutic interest as they exhibit the key characteristics and growth properties of pluripotent stem cells (8,9).
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