Western blot analysis of extracts from rat eye, mouse eye, and mouse lung using β-crystallin B1/crybb1 Rabbit Antibody (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
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
β-crystallin B1/crybb1 Rabbit Antibody recognizes endogenous levels of total β-crystallin B1 protein.Species Reactivity:
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu244 of mouse β-crystallin B1 protein. Antibodies are purified by peptide affinity chromatography.
β-crystallin B1, encoded by crybb1, is a member of the β-crystallins, one of three major crystallin types encoded by the mammalian genome, which also includes alpha- and gamma-crystallins. Crystallins are expressed at very high levels in the retinal lens and form homo/heterodimers via its N-terminal arms (1). Crystallins generally function to maintain the transparency and refractive power of the eye lens. However, they are expressed outside of the lens in other tissues and have alternative non-lens functions (2). For example, Nuc1 mutant rat, in which the βA3/A-crystallin gene is mutated, exhibit vascular remodeling abnormalities via an astrocyte-dependent mechanism (3,4). The precise cellular function of β-crystallins in glia is not well understood, but may involve regulation of endolysosomal acidification, Golgi trafficking, and specific forms of cell death (5,6). Interestingly, crybb1, is highly enriched in microglia outside of the mammalian lens and may serve as a distinct marker for microglia (7).
Explore pathways + proteins related to this product.
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.