Caspase-11 Antibody (17D9) - BSA Free

Novus Biologicals | Catalog # NB120-10454

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Bovine

Cited:

Human, Mouse, Rat, Bovine

Applications

Validated:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunoblotting, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Microarray, Knockdown Validated

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunoblotting, ELISA, Flow Cytometry, Immunoprecipitation, Proximity Ligation Assay, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG2A Clone # 17D9

Format

BSA Free
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Product Specifications

Immunogen

A recombinant p30 subunit of mouse Caspase 11. [UniProt# P70343]

Reactivity Notes

Use in Rat reported in scientific literature (PMID:32318575). Use in Bovine reported in scientific literature (PMID:31156617).

Localization

Cytoplasmic (Endoplasmic reticulum membrane and Mitochondrion)

Clonality

Monoclonal

Host

Rat

Isotype

IgG2A

Theoretical MW

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.

Scientific Data Images for Caspase-11 Antibody (17D9) - BSA Free

Western Blot: Caspase-11 Antibody (17D9)BSA Free [NB120-10454]

Western Blot: Caspase-11 Antibody (17D9)BSA Free [NB120-10454]

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 anti-rat HRP secondary antibody. Both procaspase 11 and fully cleaved caspase 11 are detected.
Immunohistochemistry-Paraffin: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454]

Immunohistochemistry-Paraffin: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454]

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 antibody generated cytoplasmic signal with more intense staining in the bronchiolar epithelium while the staining signal in alveolar cells was relatively low.
Western Blot: Caspase-11 Antibody (17D9)BSA Free [NB120-10454]

Western Blot: Caspase-11 Antibody (17D9)BSA Free [NB120-10454]

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 verified customer review.
Immunocytochemistry/ Immunofluorescence: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454]

Immunocytochemistry/ Immunofluorescence: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454]

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 (Red) and DNA (Blue) were counterstained using Phalloidin 568 and DAPI.
Western Blot: Caspase-11 Antibody (17D9)BSA Free [NB120-10454]

Western Blot: Caspase-11 Antibody (17D9)BSA Free [NB120-10454]

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 anti-Caspase 11 antibody. Image from verified customer review.
Knockout Validated: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454]

Western Blot: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454]

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 Cells were isolated from lungs, and western blot analysis was performed for pro-IL-1Beta, mature IL-1Beta, and caspase-1 cleavage. This image is from PMID: 28990935.
Knockout Validated: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454]

Western Blot: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454]

Caspase-11-Antibody-17D9-Knockout-Validated-NB120-10454-img0009.jpg
Knockout Validated: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454]

Western Blot: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454]

Caspase-11-Antibody-17D9-Knockout-Validated-NB120-10454-img0010.jpg
Knockout Validated: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454]

Western Blot: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454]

Caspase-11-Antibody-17D9-Knockout-Validated-NB120-10454-img0011.jpg
Caspase-11 Antibody (17D9) - BSA Free

Western Blot: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454] -

Western Blot: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454] - Histone induce BMECs pyroptosis is dependent on activations of caspase 1, caspase 3 & NLRP3. BMECs were incubated with stimulated histone for 16 h, & the activities of these proteins were determined by western blotting. Date are presented as mean ± SEM (n = 5). P-values of < 0.05 were considered significant (**P < 0.01, ***P < 0.001, & “ns” means not significant). Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31156617), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Caspase-11 Antibody (17D9) - BSA Free

Western Blot: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454] -

Western Blot: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454] - Caspase-11 is expressed in Casp1Null bone marrow-derived macrophages.Immunoblot analysis of caspase-1, caspase-11 & GAPDH (loading control) in unprimed WT or mutant BMDMs at various times (above lane) after stimulation with IFN-beta (250U/ml), IFN-gamma (100 ng/ml) or LPS (100 ng/ml). Data are representative of two independent experiments. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28345580), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Caspase-11 Antibody (17D9) - BSA Free

Immunohistochemistry: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454] -

Immunohistochemistry: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454] - Immunocytochemistry analysis of histone induced activations of caspase 1, caspase 3, & NLRP3 (400×). BMECs were cultured briefly on cover glasses (pre-treated with poly-L –lysine, 0.1 mg/mL, Sigma-Aldrich) & incubated with histone (50 & 100 μg/mL) for 16 h. The samples were visualized with DAB, counterstained with hematoxylin & observed by inverted microscope. Three independent experiments were carried out by light microscope analyses. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31156617), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Caspase-11 Antibody (17D9) - BSA Free

Western Blot: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454] -

Western Blot: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454] - Differential roles of caspase-1 & caspase-11 in response to infection with Aspergillus fumigatus.(A) Immunoblot analysis of pro-caspase-1 (Pro-Casp-1) & the caspase-1 subunit p20 (Casp-1 p20) & GAPDH (loading control) in unprimed WT or mutant bone marrow-derived dendritic cells left untreated (medium alone [Med]) or assessed 20 h after infection with A. fumigatus (MOI, 10). (B) Release of IL-1 beta & IL-18 after treatment as in (A). (C) Survival of 8-week-old WT & mutant mice infected with 5 × 105 A. fumigatus conidia after immunosuppression with cyclophosphamide & cortisone acetate. *P < 0.05, ****P < 0.0001 (log-rank test). Data are representative of two (C) or three independent experiments (A & B; mean & s.e.m. are representative of values from three independent experiments in B). Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28345580), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Caspase-11 Antibody (17D9) - BSA Free

Western Blot: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454] -

Western Blot: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454] - GBP2 & IRGB10 regulates caspase-11 mediated non-canonical NLRP3 activation & pyroptosis.a Microscopic analysis of cell death in unprimed WT, Gbp2−/−, Gbp5−/−, Gbpchr3-KO,Irgb10−/−, Irgb10−/−Gbpchr3-KO, Caspase11−/− & Nlrp3−/− BMDMs infected with popB (MOI 10) for 16 h. (Scale, 15 μm). Arrow heads indicate pyroptotic cells. b–e Immunoblot analysis of caspase-1, IL-1 beta, IL-18, & IL-6 release in WT, Gbp2−/−, Gbp5−/−, Gbpchr3-KO, Irgb10−/−, Irgb10−/−Gbpchr3-KO, Caspase11−/− & Nlrp3−/− BMDMs infected with popB (MOI10). f Bacterial CFU in unprimed WT, Gbp2−/−, Gbp5−/−, Gbpchr3-KO, Irgb10−/−, Irgb10−/−Gbpchr3-KO, Caspase11−/− & Nlrp3−/− BMDMs infected with popB (MOI10), 4 h & 16 h. Data are representative of three independent experiments. ns-not significant, *p < 0.05, **p < 0.01, ***p < 0.001,****p < 0.0001 (two-tailed T test) Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30062052), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Caspase-11 Antibody (17D9) - BSA Free

Flow Cytometry: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454] -

Flow Cytometry: Caspase-11 Antibody (17D9) - BSA Free [NB120-10454] - The expression of circHIPK3 in serum samples of patients with AP & in caerulein-stimulated pancreatic acinar cells. (A) QPCR was performed to detect circHIPK3 expression in serum samples of patients with AP & healthy subjects. MAP, mild acute pancreatitis; SAP, severe Acute Pancreatitis. (B) CCK8 assay was performed to measure the cell viability in AR42J cells after caerulein treatment. (C) Levels of inflammatory cytokines IL-1 beta, IL-6, IL-8, & TNF-alpha were measured by ELISA kits in culture medium after caerulein treatment. (D) Amylase activity was measured by ELISA kit in AR42J cells after caerulein treatment. (E) The pyroptosis-related proteins caspase 1 & caspase 11 were analyzed by immunoblot in AR42J cells after caerulein treatment. (F) PI staining was performed in AR42J cells after caerulein treatment, & the PI-positive cells were counted. (G) Flow-cytometric analysis of caspase-1/11+PI+ cells in AR42J cells after caerulein treatment. Data are presented as a representative plot (upper) & quantified percentages (lower). (H) circHIPK3 expression was determined by qPCR in AR42J cells after caerulein treatment. *p < 0.05. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32318575), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for Caspase-11 Antibody (17D9) - BSA Free

Application
Recommended Usage

Flow Cytometry

reported in scientific literature (PMID 27708283)

Immunoblotting

reported in scientific literature (PMID 28345580)

Immunocytochemistry/ Immunofluorescence

1:100

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Frozen

1:10-1:500

Immunohistochemistry-Paraffin

1:100

Immunoprecipitation

reported in scientific literature (PMID 10791975)

Western Blot

5 ug/ml

Reviewed Applications

Read 6 reviews rated 4 using NB120-10454 in the following applications:

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Advanced Features

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  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Caspase-11

Caspases are cysteine proteases that play an essential role in apoptosis, and 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 the lung and spleen.

Alternate Names

CASP11, CASP-11, CASP4, CASP-4, caspase 4, apoptosis-related cysteine peptidase, Caspl, ich-3

Gene Symbol

Casp4

UniProt

Additional Caspase-11 Products

Product Documents for Caspase-11 Antibody (17D9) - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for Caspase-11 Antibody (17D9) - BSA Free

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.

Citations for Caspase-11 Antibody (17D9) - BSA Free

Customer Reviews for Caspase-11 Antibody (17D9) - BSA Free (6)

4 out of 5
6 Customer Ratings
5 Stars
17%
4 Stars
67%
3 Stars
17%
2 Stars
0%
1 Stars
0%

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Customer Images


Showing  1 - 5 of 6 reviews Showing All
Filter By:
  • Caspase-11 Antibody (17D9)
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Mouse Colon
    Species: Mouse
    Verified Customer | Posted 03/31/2018
  • Caspase-11 Antibody (17D9)
    Name: Anonymous
    Application: Western Blot
    Sample Tested: mouse primary hepatocytes
    Species: Mouse
    Verified Customer | Posted 12/18/2017
    Review for anti-caspase 11 antibody (NB120-10454)
    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
    Caspase-11 Antibody (17D9) - BSA Free NB120-10454
  • Name: Anonymous
    Application: Western Blot
    Sample Tested:
    Species: Human
    Verified Customer | Posted 08/06/2016
    Review for Caspase-11 Antibody (NB120-10454)
    Caspase-11 Antibody (17D9) - BSA Free NB120-10454
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: mouse macrophage
    Species: Mouse
    Verified Customer | Posted 02/02/2016
    pro-Caspase 11 western blot
    Caspase-11 Antibody (17D9) - BSA Free NB120-10454
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: Bone Marrrow Derived Macrophages
    Species: Mouse
    Verified Customer | Posted 03/19/2015
    Caspase 11 with LPS Stimulation
    Caspase-11 Antibody (17D9) - BSA Free NB120-10454
  • Name: Hyun Yong Jin
    Application: Western Blot
    Sample Tested:
    Species: Mouse
    Verified Customer | Posted 07/01/2014
    Mouse B cell
    Caspase-11 Antibody (17D9) - BSA Free NB120-10454

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Showing  1 - 5 of 6 reviews Showing All

Protocols

View specific protocols for Caspase-11 Antibody (17D9) - BSA Free (NB120-10454):

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and wash the cells briefly in PBS. Add 10% formalin to the dish and fix at room temperature for 10 minutes.
2. Remove the formalin and wash the cells in PBS.
3. Permeablize the cells with 0.1% Triton X100 or other suitable detergent for 10 min.
4. Remove the permeablization buffer and wash three times for 10 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 10 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 10 minutes each.
10. Counter stain DNA with DAPi if required.

Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in 1% BSA and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Caspase-11 Antibody (17D9) - BSA Free

Showing  1 - 2 of 2 FAQs Showing All
  • Q: 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).

    A: 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.

  • Q: 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?

    A: 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.

  • Q: 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).

    A: 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.

  • Q: 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?

    A: 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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