Caspase-11 Antibody (17D9) - BSA Free

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) [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) [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) [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) [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) [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) [NB120-10454] - CNF1-triggered immunity requires inflammatory caspases.(A and B) IL-1beta production and maturation/secretion after treatment with CNF1, LPS or CNF1 (1 ug/ml) + LPS (100 ng/ml) for 10 h. Actin and BSA were used as loading controls. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.ppat.1004732), licensed under a CC-BY license.

Knockout Validated: Caspase-11 Antibody (17D9) [NB120-10454] - CNF1-triggered IL-1beta maturation requires activated Rac, ASC and caspase-1. Western blot analysis of the production and maturation/secretion of IL-1beta by primary macrophages following treatment with CNF1, LPS or CNF1+LPS for 10 h. Actin and BSA were used as loading controls. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.ppat.1004732), licensed under a CC-BY license.

Knockout Validated: Caspase-11 Antibody (17D9) [NB120-10454] - Caspase-11 is expressed in Casp1Null bone marrow-derived macrophages. Immunoblot analysis of caspase-1, caspase-11 and GAPDH (loading control) in unprimed WT 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 and cropped by CiteAb from the following publication (//www.nature.com/articles/srep45126), licensed under a CC-BY license.

• Reactivity: Hu, Mu, Rt, Bv
• Applications: WB, ELISA, Flow, ICC/IF, IHC, IP, MA, WB, KD, KO
• Clone: 17D9
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

Caspase-11 Antibody (17D9) - BSA Free Summary

• Immunogen: A recombinant p30 subunit of mouse Caspase 11. [UniProt# P70343]
• Localization: Cytoplasmic (Endoplasmic reticulum membrane and Mitochondrion)
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Rat
• Gene: Casp4
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • ELISA
  • 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)
  • Knockdown Validated
  • Knockout Validated
  • Microarray
  • Western Blot 5 ug/ml
• 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.
• Control:

  Jurkat Staurosporine Treated / Untreated Cell Lysate

Reactivity Notes

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

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for Caspase-11 Antibody (17D9) - BSA Free

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

Background

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.

Limitations

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.

Publications for Caspase-11 Antibody (NB120-10454)(67)

Publications using NB120-10454

• Lin A, Guo Y, Zhang H et al. GSDMD knockdown exacerbates hippocampal damage and seizure susceptibility by crosstalk between pyroptosis and apoptosis in kainic acid-induced temporal lobe epilepsy Biochimica et biophysica acta. Molecular basis of disease 2023-03-14 [PMID: 36921736]
• Han J, Tweedell R, Kanneganti T Evaluation of Caspase Activation to Assess Innate Immune Cell Death Journal of Visualized Experiments 2023-01-20 [PMID: 36744800]
• Saxena P Role of Inflammatory Cytokine Signaling in Control of Bacterial Infection Thesis 2020-01-01 (WB, Mouse)
• Eren E, Planes R, Buyck JM et al. Type-3 Secretion Systeminduced pyroptosis protects Pseudomonas against cell-autonomous immunity bioRxiv (WB, Mouse)
• Daily KP, Badr A, Eltobgy M et al. DNA hypomethylation promotes the expression of CASPASE-4 which exacerbates neuroinflammation and amyloid-? deposition in Alzheimer's disease The Ohio State University College of Medicine bioRxiv 2023-09-01 [PMID: 37693600] (WB)
• Exconde PM, Hernandez-Chavez C, Bray MB et al. The tetrapeptide sequence of IL-1? regulates its recruitment and activation by inflammatory caspases bioRxiv : the preprint server for biology 2023-02-16 [PMID: 36824844] (WB, Human)
• Wu YH, Mo ST, Chen IT et al. Caspase-8 inactivation drives autophagy-dependent inflammasome activation in myeloid cells Science advances 2022-11-11 [PMID: 36367942] (WB, Mouse)
• Gros M, Segura E, Rookhuizen DC et al. Endocytic membrane repair by ESCRT-III controls antigen export to the cytosol during antigen cross-presentation Cell reports 2022-08-16 [PMID: 35977488] (WB)
• Jiang H, Niu C, Guo Y et al. Wedelolactone induces apoptosis and pyroptosis in retinoblastoma through promoting ROS generation International immunopharmacology 2022-07-26 [PMID: 35905560]
• Lin WY, Li LH, Hsiao YY et al. Repositioning of the Angiotensin II Receptor Antagonist Candesartan as an Anti-Inflammatory Agent With NLRP3 Inflammasome Inhibitory Activity Frontiers in Immunology 2022-05-20 [PMID: 35669789] (B/N)
• Wang Z, Yang X, Wang X et al. Glycyrrhizin attenuates caspase-11-dependent immune responses and coagulopathy by targeting high mobility group box 1 International immunopharmacology 2022-03-24 [PMID: 35339905] (PLA, WB, IF/IHC, Mouse, Human)
• Sundaram B, Karki R, Kanneganti TD NLRC4 Deficiency Leads to Enhanced Phosphorylation of MLKL and Necroptosis ImmunoHorizons 2022-03-17 [PMID: 35301258] (WB, Mouse)
• Huang D, Chen P, Huang G et al. Salt-inducible kinases inhibitor HG-9-91-01 targets RIPK3 kinase activity to alleviate necroptosis-mediated inflammatory injury Cell death & disease 2022-02-25 [PMID: 35217652] (WB, Mouse)
• Guo Y, Mao R, Xie Q et al. Francisella novicida Mutant XWK4 Triggers Robust Inflammasome Activation Favoring Infection Frontiers in Cell and Developmental Biology 2021-11-18 [PMID: 34869331] (WB)
• Zheng M, Karki R, Kancharana B Et al. Caspase-6 promotes activation of the caspase-11-NLRP3 inflammasome during gram-negative bacterial infections Journal of Biological Chemistry 2021-11-01 [PMID: 34740613] (WB)
• Karki R, Sundaram B, Sharma BR Et al. ADAR1 restricts ZBP1-mediated immune response and PANoptosis to promote tumorigenesis Cell reports 2021-10-19 [PMID: 34686350] (WB, Mouse)
• Xiong S, Zhang L, Richner JM et al. Interleukin-1RA Mitigates SARS-CoV-2-Induced Inflammatory Lung Vascular Leakage and Mortality in Humanized K18-hACE-2 Mice Arteriosclerosis, thrombosis, and vascular biology 2021-09-09 [PMID: 34496633]
• Fan R, Sui J, Dong X et al. Wedelolactone alleviates acute pancreatitis and associated lung injury via GPX4 mediated suppression of pyroptosis and ferroptosis Free radical biology & medicine 2021-07-08 [PMID: 34246777] (WB)
• Wang X, Li Z, Bai Y et al. A small molecule binding HMGB1 inhibits caspase-11-mediated lethality in sepsis Cell death & disease 2021-04-14 [PMID: 33854044]
• Chen Q, Zheng J, Wang D, et al. Nitrosonisoldipine is a selective inhibitor of inflammatory caspases and protects against pyroptosis and related septic shock European journal of immunology 2021-01-17 [PMID: 33454984] (WB, Mouse)
• Karki R, Sharma BR, Tuladhar S et al. Synergism of TNF- alpha and IFN-gamma Triggers Inflammatory Cell Death, Tissue Damage, and Mortality in SARS-CoV-2 Infection and Cytokine Shock Syndromes Cell 2020-11-19 [PMID: 33278357] (WB, Mouse)
• Karki R, Sharma BR, Tuladhar S et al. Synergism of TNF-? and IFN-? triggers inflammatory cell death, tissue damage, and mortality in SARS-CoV-2 infection and cytokine shock syndromes bioRxiv 2023-02-09 [PMID: 33140051] (WB)
• Finethy R, Dockterman J, Kutsch M, Orench-Rivera N Dynamin-related Irgm proteins modulate LPS-induced caspase-4 activation and septic shock EMBO Rep 2020-10-30 [PMID: 33124745]
• Sakaguchi N, Sasai M, Bando H et al. Role of Gate-16 and Gabarap in Prevention of Caspase-11-Dependent Excess Inflammation and Lethal Endotoxic Shock Frontiers in Immunology 2020-09-15 [PMID: 33042141] (ICC/IF, WB)
• Wang Y, Xiao Y, Long S et al. Gasdermin D in macrophages restrains colitis by controlling cGAS-mediated inflammation Sci Adv 2020-07-17 [PMID: 32671214] (WB, Mouse)
• Selkrig J, Li N, Hausmann A et al. Spatiotemporal proteomics uncovers cathepsin-dependent macrophage cell death during Salmonella infection Nat Microbiol 2020-06-08 [PMID: 32514074] (WB, Mouse)
• Wang J, Li X, Liu Y et al. CircHIPK3 Promotes Pyroptosis in Acinar Cells Through Regulation of the miR-193a-5p/GSDMD Axis Front Med (Lausanne) 2020-04-07 [PMID: 32318575] (FLOW, Rat)
• Karki R, Lee E, Sharma Br et Al. IRF8 Regulates Gram-Negative Bacteria-Mediated NLRP3 Inflammasome Activation and Cell Death J. Immunol. 2020-03-23 [PMID: 32205422] (IB, Mouse)
• Huang LS, Hong Z, Wu W et al. mtDNA Activates cGAS Signaling and Suppresses the YAP-Mediated Endothelial Cell Proliferation Program to Promote Inflammatory Injury Immunity 2020-03-17 [PMID: 32164878]
• Wang C, Li Y, Wu X et Al. IRF3 and IRF7 contribute to diesel exhaust particles-induced pulmonary inflammation by mediating mTORC1 activation and restraining autophagy in mice Eur. J. Immunol. 2020-03-05 [PMID: 32135578] (WB, Mouse)
• Wang K, Sun Q, Zhong X et al. Structural Mechanism for GSDMD Targeting by Autoprocessed Caspases in Pyroptosis Cell 2020-02-17 [PMID: 32109412] (WB, Mouse, Human)
• Li S, Wu Y, Yang D et al. Gasdermin D in peripheral myeloid cells drives neuroinflammation in experimental autoimmune encephalomyelitis J. Exp. Med. 2019-08-29 [PMID: 31467036] (IB, Mouse)
• Wei Z, Wang J et al. Effects of Neutrophil Extracellular Traps on Bovine Mammary Epithelial Cells in vitro. Front Immunol 2019-04-06 [PMID: 31156617] (WB, Bovine)
• Kayagaki N, Lee BL, Stowe IB et al. IRF2 transcriptionally induces GSDMD expression for pyroptosis Sci Signal 2019-05-21 [PMID: 31113851]
• Pronin A, Pham D, An W et al. Inflammasome Activation Induces Pyroptosis in the Retina Exposed to Ocular Hypertension Injury Front Mol Neurosci 2019-03-13 [PMID: 30930743] (IHC-Fr, Mouse)
• Liu Q, Zhang S, Sun Z et al. E3 ubiquitin ligase Nedd4 is a key negative regulator for non-canonical inflammasome activation Cell Death Differ. 2019-02-28 [PMID: 30816303] (IHC-P, Human)
• Wang B, Lim JH, Kajikawa T et al. Macrophage b2-Integrins Regulate IL-22 by ILC3s and Protect from Lethal Citrobacter rodentium-Induced Colitis Cell Rep 2019-02-05 [PMID: 30726742] (WB, Mouse)
• Quach J, Moreau F, Sandall C, Chadee k. Entamoeba histolytica-induced IL-1B secretion is dependent on caspase-4 and gasdermin D. Mucosal Immunol. 2018-10-25 [PMID: 30361535] (IP, Human)
• Chen Jing, Wang Bei, Lai Jinsheng et al. Trimetazidine Attenuates Cardiac Dysfunction in Endotoxemia and Sepsis by Promoting Neutrophil Migration. Frontiers in Physiology 2018-09-04 [PMID: 30233596] (WB, IF/IHC, Mouse)
• Balakrishnan A, karki R, Berwin B et al. Guanylate binding proteins facilitate caspase-11-dependent pyroptosis in response to type 3 secretion system-negative Pseudomonas aeruginosa. Cell Death Discov. 2018-06-27 [PMID: 30062052] (WB, Mouse)
• Deigendesch N, Zychlinsky A, Meissner F. Copper Regulates the Canonical NLRP3 Inflammasome. J. Immunol. 2018-03-01 [PMID: 29358279] (Mouse)
• Kerur N, Fukuda S, Banerjee D et al. cGAS drives noncanonical-inflammasome activation in age-related macular degeneration Nat. Med. 2017-11-27 [PMID: 29176737] (Human)
• Cheng KT, Xiong S, Ye Z et al. Caspase-11-mediated endothelial pyroptosis underlies endotoxemia-induced lung injury. J. Clin. Invest. 2017-11-01 [PMID: 28990935] (WB, Mouse)
• Ming Man S, Karki R, Briard B et al. Differential roles of caspase-1 and caspase-11 in infection and inflammation. Sci Rep. 2017-03-27 [PMID: 28345580] (IB, Mouse)
• Jin HY, Oda H, Chen P et al. Differential Sensitivity of Target Genes to Translational Repression by miR-17-92. PLoS Genet. [PMID: 28241004] (WB, Mouse)
• Klaska IP, Muckersie E, Martin-Granados C et al. LPS-primed Heterotolerant Dendritic Cells suppress Experimental Autoimmune Uveoretintitis by multiple mechanisms. Immunology. 2016-11-15 [PMID: 27859049] (WB, Mouse)
• Kolb Ryan, Phan Liem, Borcherding Nicholas et al. Obesity-associated NLRC4 inflammasome activation drives breast cancer progression. Nat Commun 2016-01-01 [PMID: 27708283] (FLOW, Mouse)
• Man S, Karki R, Sasai M et al. IRGB10 Liberates Bacterial Ligands for Sensing by the AIM2 and Caspase-11-NLRP3 Inflammasomes. Cell 2016-09-29 [PMID: 27693356] (WB, Mouse)
• Kim MJ, Bae SH, Ryu JC et al. SESN2/sestrin2 suppresses sepsis by inducing mitophagy and inhibiting NLRP3 activation in macrophages. Autophagy 2016-06-23 [PMID: 27337507] (IP, WB)
• Chung H, Vilaysane A, Lau A et al. NLRP3 regulates a non-canonical platform for caspase-8 activation during epithelial cell apoptosis. Cell Death Differ. 2016-02-19 [PMID: 26891693] (WB, Mouse)
• Kayagaki N, Stowe IB, Lee BL et al. Caspase-11 cleaves gasdermin D for non-canonical inflammasome signaling. Nature 2015-09-16 [PMID: 26375259] (WB, Mouse)
• Shi J, Zhao Y, Wang K et al. Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death. Nature 2015-09-16 [PMID: 26375003] (WB, Mouse)
• Aachoui Y, Kajiwara Y, Leaf IA et al. Canonical Inflammasomes Drive IFN-gamma to Prime Caspase-11 in Defense against a Cytosol-Invasive Bacterium. Cell Host Microbe 2015-09-09 [PMID: 26320999] (WB, Mouse)
• Shaw D, Wang X, Brown L et al. Infection-derived lipids elicit an immune deficiency circuit in arthropods. Nature Communications. [PMID: 25790189]
• Diabate M, Munro P, Garcia E et al. Escherichia coli a-Hemolysin Counteracts the Anti-Virulence Innate Immune Response Triggered by the Rho GTPase Activating Toxin CNF1 during Bacteremia. PLoS Pathog. 2015-03-01 [PMID: 25781937] (ELISA, Mouse)
• Youm YH, Nguyen KY, Grant RW et al. The ketone metabolite beta-hydroxybutyrate blocks NLRP3 inflammasome-mediated inflammatory disease Nat. Med. 2015-02-16 [PMID: 25686106] (WB, Human)
• Demon D, Kuchmiy A, Fossoul A et al. Caspase-11 is expressed in the colonic mucosa and protects against dextran sodium sulfate-induced colitis. Mucosal Immunology 2014-01-01 [PMID: 24850431] (WB, Mouse)
• Yuan Z, Syed MA, Panchal D et al. TREM-1 mediated Bcl-2 induction prolongs macrophage survival. J. Biol. Chem. 2014-04-07 [PMID: 24711453] (WB, Mouse)
• Van Opdenbosch Nina, Gurung Prajwal, Vande Walle Lieselotte et al. Activation of the NLRP1b inflammasome independently of ASC-mediated caspase-1 autoproteolysis and speck formation. Nat Commun 2014-01-01 [PMID: 24492532] (WB)
  
  Details:
  Using the HRP conjugated version of NB600-501, catalog number NB600-501H.
• Gurung P, Anand PK, Malireddi RK et al. FADD and Caspase-8 Mediate Priming and Activation of the Canonical and Noncanonical Nlrp3 Inflammasomes. J. Immunol. 2014-02-10 [PMID: 24453255] (WB, Mouse)
• Jin HY, Oda H, Lai M et al. MicroRNA-17-92 plays a causative role in lymphomagenesis by coordinating multiple oncogenic pathways. EMBO J. 2013-08-28 [PMID: 23921550] (WB, Mouse)
• Case CL, Kohler LJ, Lima JB et al. Caspase-11 stimulates rapid flagellin-independent pyroptosis in response to Legionella pneumophila Proc Natl Acad Sci U S A 2013-01-10 [PMID: 23307811] (WB, Mouse)
• Sun L, Wu Z, Hayashi Y et al. Microglial Cathepsin B Contributes to the Initiation of Peripheral Inflammation-Induced Chronic Pain J Neurosci 2012-08-15 [PMID: 22895716] (WB, Mouse)
• Kenneth NS, Younger JM, Hughes ED et al. An inactivating caspase 11 passenger mutation originating from the 129 murine strain in mice targeted for c-IAP1 Biochem J 2012-04-15 [PMID: 22332634] (WB, Mouse)
• Kayagaki N, Warming S, Lamkanfi M et al. Non-canonical inflammasome activation targets caspase-11. Nature. 2011-10-16 [PMID: 22002608]
• Kang SJ, Wang S, Kuida K, Yuan J. Distinct downstream pathways of caspase-11 in regulating apoptosis and cytokine maturation during septic shock response. Cell Death Differ. 2002-10-01 [PMID: 12232800] (IF/IHC, Mouse)
• Kang SJ, Wang S, Hara H et al. Dual role of caspase-11 in mediating activation of caspase-1 and caspase-3 under pathological conditions. J Cell Biol. 2000-05-01 [PMID: 10791975] (WB, IP, IF/IHC, Mouse)

Reviews for Caspase-11 Antibody (NB120-10454) (6)

Average Rating: 4

(Based on 6 reviews)

We have 6 reviews tested in 2 species: Human, Mouse.

reviewed by: Verified Customer WB Mouse 03/31/2018

Summary

• Application: Western Blot
• Sample Tested: Mouse Colon
• Species: Mouse
• Lot: A-10

reviewed by: Verified Customer WB Mouse 12/18/2017

Summary

• Application: Western Blot
• Sample Tested: mouse primary hepatocytes
• Species: Mouse
• Lot: A-8

Comments

• Comments: 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

reviewed by: Verified Customer WB Human 08/06/2016

Summary

• Application: Western Blot
• Species: Human
• Lot: PO number: P00129026

reviewed by: Verified Customer WB Mouse 02/02/2016

Summary

• Application: Western Blot
• Sample Tested: mouse macrophage
• Species: Mouse
• Lot: A-6

reviewed by: Verified Customer WB Mouse 03/19/2015

Summary

• Application: Western Blot
• Sample Tested: Bone Marrrow Derived Macrophages
• Species: Mouse
• Lot: 102M4832

reviewed by: Hyun Yong Jin WB Mouse 07/01/2014

Summary

• Application: Western Blot
• Species: Mouse

FAQs for Caspase-11 Antibody (NB120-10454). (Showing 1 - 2 of 2 FAQs).

1. 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.
2. 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.

Bioinformatics

• Gene Symbol: Casp4
• Uniprot:
  • P70343()