|H M R Mk||Endogenous||21||Rabbit|
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
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.
FTH1 Antibody detects endogenous levels of total FTH1 protein. Nonspecific bands are seen above 80 kDa.
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino-terminal region of FTH1. Antibodies are purified by protein A and peptide affinity chromatography.
Ferritin (FTH) is a ubiquitous and highly conserved protein which plays a major role in iron homeostasis by sequestering and storing iron in a non-toxic and bioavailable form (1). The assembled ferritin molecule, often referred to as a nanocage, can store up to 4,500 atoms of iron (2,3). It forms a holoenzyme of ~450 kDa, consisting of 24 subunits made up of two types of polypeptide chains: ferritin heavy chain and ferritin light chain, each having unique functions. Ferritin heavy chains catalyze the first step in iron storage, the oxidation of Fe(II), whereas ferritin light chains promote the nucleation of ferrihydrite, enabling storage of Fe(III) (4). In addition to iron buffering, heavy chain ferritin also enhances thymidine biosynthesis (5). Serum ferritin levels serve as an indicator of the amount of iron stored in the body. Serum ferritin is the most sensitive test for anaemia. The level of serum ferritin is markedly elevated in inflammation, malignancy, and iron overload disorders (6). Research studies have found that defects in ferritin proteins are also associated with several neurodegenerative diseases (7).
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