Biological Strategies: Western Blot: Caspase-11 Antibody (17D9) [NB120-10454] - Analysis of Caspase-11 in THP-1 cell lysates that were untreated (left lane) or treated (right lane) with 100 ng/ml LPS. Image from ...read more
Genetic Strategies: Western Blot: Caspase-11 Antibody (17D9) [NB120-10454] - Casp11fl/fl and Casp11EC-/- mice (n = 3) were challenged with LPS (40 mg/kg i.p.) for 6 hours. Endothelial Cellss were isolated from ...read more
Immunocytochemistry/ Immunofluorescence: Caspase 11 Antibody (17D9) [NB120-10454] - Caspase 11 antibody was tested in NIH-3T3 cells at a 1:100 dilution using a Dylight 488 conjugated secondary antibody (Green). Actin ...read more
Immunohistochemistry-Paraffin: Caspase 11 Antibody (17D9) [NB120-10454] - IHC-P analysis of a formalin fixed paraffin embedded tissue section of mouse lung using Caspase 11 antibody (clone 17D9) at 1:100 diluton. The ...read more
Western Blot: Caspase 11 Antibody (17D9) [NB120-10454] - Mouse B cell. Image from verified customer review.
Western Blot: Caspase 11 Antibody (17D9) [NB120-10454] - Mouse liver extract was separated on a 4-15% gel and transfered to PVDF membrane. The membrane was probed with anti-caspase 11 antibody and detected with an ...read more
Biological Strategies: Western Blot: Caspase-11 Antibody (17D9) [NB120-10454] - Caspase 11 Antibody (17D9) [NB120-10454] - analysis of Caspase 11 in LPS stimulated bone marrow derived macrophages using ...read more
Use in immunprecipitation reported in scientific literature (PMID 10791975). Use in immunoblotting reported in scientific literature (PMID: 28345580). The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
35-43 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Read 6 Reviews rated 4 using NB120-10454 in the following applications:
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
0.02% Sodium Azide
Protein G purified
Alternate Names for Caspase-11 Antibody (17D9)
caspase 4, apoptosis-related cysteine peptidase
Caspases are cysteine proteases that play an essential role in apoptosis; their regulation is post-translational to enable rapid activation. Murine Caspase 11, also known as Caspase 4 or ICH3, has a heterotetrameric structure consisting of two anti-parallel heterodimers, each one formed of a large (p20) and a small (p10) subunit. Each heterodimer has a weight of 43kDa. Upon activation, murine caspase 11 is cleaved by an autocatalytic mechanism to give rise to the individual subunits. Expression levels of murine Caspase 11 are highest in lung and spleen.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
100 ng/ml LPS treated for 1 hour Name: Anti-caspase 11 antibody (NB120-10454) Catalog #: Anti-caspase 11 antibody (NB120-10454) Lot Number: Anti-caspase 11 antibody (NB120-10454, Lot # A-6) PO/Order Number: Click here to enter text.. WB Image Description (Please provide labels for all lanes): mouse primary hepatocytes treated without (lane 1), or with 100 ng/ml LPS (lane 2) Sample Information: Cell Line or Tissue: mouse primary hepatocytes Species: mouse Treatment: 100 ng/ml LPS Lysate Preparation: Date of lysate preparation: December 4, 2017 Lysis buffer used: 1X lysis buffer from Cell Signaling by adding PMSF Reducing agent: beta-mercaptoethanol, DTT If boiled (temperature/time): Yes Controls: Positive Control: No Negative Control: No Loading Control (please attach additional images if applicable): No Protein Amount Loaded per lane: 20 ug Antibody Storage Conditions: -20℃ Electrophoresis: Gel Percentage: 12% Electrophoresis Conditions: Tris-Glycine-SDS at room temperature Voltage: 120V Time: 2 hours Membrane Transfer: Method (Submersion/Semi-dry): wet transfer Membrane Type (PVDF/Nitrocellulose): Nitrocellulose Time: 2 hours Voltage: 100V Blocking: Blocking Solution: 5% milk in 1X TBST Time: 1 hour at room temperature Primary Antibody: Dilution: 1/200 Diluent Buffer: 2.5% BSA Incubation Time: overnight Incubation Temperature: 4℃ Washing Conditions: Wash Solution: 1X TBST Time and Repetitions: 5 min each for 3 times Secondary Antibody Manufacturer and Catalog #: Thermo Scientific, cat # 34095, Lot # SG251180 Secondary description: goat anti-rat secondary antibody Dilution: 1/5000 Diluent Buffer: 3% milk Incubation Time: 1 hour Incubation Temperature: room temperature Detection Method: Detection: ECL (GE, cat # RPN2209, lot #n9838243) Procedure: Add equal volume of A and B, mix and apply on the membranes for 3-5 min before exposure Development Time: 25 seconds Molecular weight of band(s): ~ 38, 43 kDa Experimental Concerns and Observations: Specific bands around 38, 43 kDa were observed
this is a very good antibody for pro-Caspase 11 detection, its specificity against the activated form of caspase 11 remains unknown, at least we didn't see a strong potency for the detection of activated form through western blot.
FAQs for Caspase-11 Antibody (NB120-10454). (Showing 1 - 2 of 2 FAQs).
If I dilute the 100 uL of antibody (the whole delivered content) in 10 mL of buffer to incubate my blottet membrane in (and I need this volume of about 10 mL for the membrane to float in it on the shaker). I will be able to re-use this one dilution (which is 10 ug/mL) for 100 western blots? So that means for 100 samples? Or even for 100 membranes? Usually we use a antibody-dilution of 1:1000 (around 1ug/mL), this would be 1:100 (10ug/mL).
I am hoping I can assist you in determining usage of this antibody in your assay. The antibody comes at a starting concentration of 1 mg/mL. Since you get 100uL in the vial you will have 100 ug of antibody. The lab recommends using a dilution of 1:100-1:200 for WB. Therefore if you wished to use 10 mL volume on your incubation you would need to add 100 uL of antibody into 9.9mL of buffer to give you a final concentration of 10 ug/uL. Since you are using such a large final volume you will only get 1-2 blots per Western ran since we do not recommend reusing your antibody. If you can run the blots with much lower final volume such as 1 mL instead you can get away with a lot more Westerns.
If it has been determined, can you let me know as specifically as possible the epitope that the Caspase-11 antibody recognizes, eg. peptide sequence or approximate region of the p30 subunit?
This antibody has not been epitope mapped and was generated using full-length p30. The only information we have is that the antibody may be used for immunoblotting [to detect the full length caspase 11 (apparent size of 35-43 kDa), and the p10 and p30fragments], so it must recognize a region similar in both subunits.
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