REACTIVITY | SENSITIVITY | MW (kDa) | Isotype |
---|---|---|---|
H M | Endogenous | 17,31 | Mouse IgG1 |
Western blot analysis of extracts from THP-1 cells, untreated (-) or LPS-treated (100 ng/mL, 3 hr; +), using IL-1β (3A6) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Learn more about how we get our imagesWestern blot analysis of extracts from Raw 264.7 cells, untreated (-) or treated with Brefeldin A #9972 (300 ng/mL, last 3 hr of stimulation; +) and LPS (100 ng/mL, 6 hr; +), using IL-1β (3A6) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Learn more about how we get our imagesWestern blot analysis of 1 ng recombinant Human Interleukin-1β (hIL-1β) #8900 and 1 ng recombinant Mouse Interleukin-1β (mIL-1β) #5204 using IL-1β (3A6) Mouse mAb.
Learn more about how we get our imagesImmunohistochemical analysis of paraffin-embedded human large intestine (chronic colitis of the colon) using IL-1-β (3A6) Mouse mAb.
Learn more about how we get our imagesImmunohistochemical analysis of paraffin-embedded human large intestine (ulcerative chronic colitis of the rectum) using IL-1-β (3A6) Mouse mAb.
Learn more about how we get our imagesImmunohistochemical analysis of paraffin embedded THP-1 cell pellets, control (left) or LPS-treated (right) using IL-1-β (3A6) Mouse mAb.
Learn more about how we get our imagesFor 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.
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: Loading of prestained molecular weight markers (#13953, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended.
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: 262
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Do not allow slides to dry at any time during this procedure.
For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; follow with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.
posted February 2010
revised March 2016
Protocol Id: 280
Application | Dilutions |
---|---|
Western Blotting | 1:1000 |
Immunohistochemistry (Paraffin) | 1:100 |
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 (3A6) Mouse mAb recognizes endogenous levels of total IL-1β protein.
Human, Mouse
Monoclonal antibody is produced by immunizing animals with recombinant human 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.
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Product # | Size | Price |
---|---|---|
12242S | 100 µl (10 western blots) | $ 255.0 |