|M||Endogenous||17, 31||Rabbit IgG|
Western blot analysis of extracts from Raw264.7 cells, untreated (-) or treated with LPS (100 ng/mL, 6 hr; +), using IL-1β (D6D6T) Rabbit mAb (Mouse Specific) (upper), or β-Actin (D6A8) Rabbit mAb #8457 (lower).Learn more about how we get our images.
Western blot analysis of recombinant mouse IL-1β (mIL-1β) using IL-1β (D6D6T) Rabbit mAb (Mouse Specific).Learn more about how we get our images.
Flow cytometric analysis of Raw264.7 cells, untreated (blue) or treated with LPS (100 ng/ml, 6 hr) (green) using IL-1β (D6L6T) Rabbit mAb (Mouse Specific). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.Learn more about how we get our images.
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
NOTE: Prepare solutions with RODI (reverse osmosis deionized) or equivalent grade water.
NOTE: Immunostaining of surface antigens should be performed on live cells prior to fixation/permeabilization, as per manufacturer’s requirements.
posted December 2013
Protocol Id: 626
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.
IL-1β (D6D6T) Rabbit mAb (Mouse Specific) recognizes endogenous levels of total IL-1β protein.
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to mouse IL-1β protein.
Interleukin-1β (IL-1β), one of the major caspase-1 targets, is a multifunctional cytokine that is involved in a host of immune and proinflammatory responses (1). It is produced primarily by activated monocytes and macrophages. It signals through various adaptor proteins and kinases that lead to activation of numerous downstream targets (2-6). Human IL-1β is synthesized as a 31 kDa precursor. To gain activity, the precursor must be cleaved by caspase-1 between Asp116 and Ala117 to yield a 17 kDa mature form (7,8). Detection of the 17 kDa mature form of IL-1β is a good indicator of caspase-1 activity.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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|31202S||100 µl (10 western blots)||$ 255.0|