|H M R||Endogenous||35-45||Rabbit IgG|
Western blot analysis of extracts from various cell lines using hnRNP E2/PCBP2 (D1S5E) Rabbit mAb (upper) or Vinculin (E1E9V) XP® Rabbit mAb (HRP Conjugate) #18799 (lower).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: 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
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.
hnRNP E2/PCBP2 (D1S5E) Rabbit mAb recognizes endogenous levels of total hnRNP E2/PCBP2 protein.
Human, Mouse, Rat
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu243 of human hnRNP E2/PCBP2 protein.
PCPB2 (also known as hnRNP E2) is an RNA-binding protein that interacts in a sequence-specific fashion with single-stranded poly (rC). Through their poly(rC)-binding ability, PCBPs regulate mRNA stability and translation (1,2). PCBP2 is an iron chaperone; it delivers iron to ferritin for storage and mediates metalation of certain iron-containing proteins (3-5). PCBP2 interacts with the iron importer DMT1 (divalent metal transporter 1) and the iron exporter FPN1 (ferroportin 1) and regulates their activities (6,7). PCBP2 is induced by viral infection and targets MAVS for polyubiquitination and degradation (8-10). Recent reports demonstrate that it is involved in Hippo signaling, miRNA processing, immune suppression, and cancer (11-15).
Cell Signaling Technology 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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|83017S||100 µl||$ 255.0|