Western blot analysis of extracts from various cell lines using KIF5B Antibody (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from HCT 116 and HCT 116 KIF5B knockout (-/-) cells using KIF5B Antibody (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
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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
KIF5B Antibody recognizes endogenous levels of total KIF5B protein.Species Reactivity:
Human, Mouse, Rat
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys32 of human KIF5B protein. Antibodies are purified by peptide affinity chromatography.
Kinesin superfamily proteins (KIFs) are molecular motors that drive directional, microtubule-dependent intracellular transport of membrane-bound organelles and other macromolecules (e.g. proteins, nucleic acids). The intracellular transport functions of KIFs are fundamentally important for a variety of cellular functions, including mitotic and meiotic division, motility/migration, hormone and neurotransmitter release, and differentiation (1-4). Disruptions to KIF-mediated intracellular transport have been linked with a variety of pathologies, ranging from tumorigenesis to defects in higher order brain function such as learning and memory (4-6).
KIF5 consists of three family members referred to as KIF5A, KIF5B, and KIF5C (7). KIF5A and KIF5C are specifically expressed in neurons whereas KIF5B is expressed ubiquitously (7-10). Targeted disruption of the kif5B gene resulted in embryonic lethality (11). These and other studies demonstrate that KIF5B plays an important role in the localization and distribution of major organelles, including the mitochondria and lysosome, and can contribute to autophagy (11-13). In addition, gene rearrangements involving KIF5B fusions to ALK and RET have been identified as drivers for lung cancer and other malignancies (14-18).
Explore pathways + proteins related to this product.
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