Natural Killer (NK) And Natural Killer T (NKT) Cells
NK and NKT CellsNatural Killer (NK)and Natural Killer T (NKT) CellsTools for Research and ResultsTopicsNK Cell ReceptorsLy-49, KIR and NCR FamiliesNK Cell Activation and FunctionActivate, Stimulate and ProliferateIntracellular Flow Cytometry StainingImmunoassay QuantitationNK Cell Transcriptional ControlNK T CellsNK Cell Marker GuideeBioscience (US) Tel: 1-888-999-1371Affymetrix, Inc. Tel: 1-888-362-2447Tel: 1-858-642-2058eBioscience (EU) Tel: 43 1 796 40 40 304Affymetrix UK Ltd. Tel: 44-(0)1628-552550Panomics Solutions Tel: 1-877-726-6642 rix Japan K.K. Tel: 81-(0)3-6430-4020USB Products Tel: 1-800-321-9322 usb.affymetrix.comwww.ebioscience.com Please visit our website for international distributor contact information.For Research Use Only. Not for use in diagnostic or therapeutic procedures.IMM02507-1 NK and NKT Reagents PLF 0913 Affymetrix, Inc. All rights reserved. Affymetrix , Axiom , Command Console , CytoScan , DMET , GeneAtlas , GeneChip , GeneChip-compatible , GeneTitan , Genotyping Console ,myDesign , NetAffx , OncoScan , Powered by Affymetrix , PrimeView , Procarta and QuantiGene are trademarks or registered trademarks of Affymetrix, Inc. All other trademarks are theproperty of their respective owners. BestProtocols , eBioscience , eFluor , Full Spectrum Cell Analysis , InstantOne ELISA , OneComp eBeads , ProcartaPlex , Ready-SET-Go! , SAFE SuperAquaBlue , The New Standard of Excellence and UltraComp eBeads are trademarks or registered trademarks of eBioscience, Inc. Instant ELISA is a registered trademark of Bender MedSystems,GmbH. FlowLogic is a registered trademark of Inivai TechnologiesProducts may be covered by one or more of the following patents: U.S. Patent Nos. 5,445,934; 5,744,305; 5,945,334; 6,140,044; 6,399,365; 6,420,169; 6,551,817; 6,733,977; 7,629,164;7,790,389 and D430,024 and other U.S. or foreign patents. Products are manufactured and sold under license from OGT under 5,700,637 and 6,054,270. Cyanine (Cy) dye conjugates are coveredby US Patent Nos. US5,569,587 and US5,627,027.Natural killer (NK) cells are lymphoid cells poised and ready toassist in the destruction of virally infected cells and tumor cellsfrom the body. NK cells are part of the innate immune systemand mediate their effect in an antigen-independent mannerthat, in general, does not give rise to immunological memoryor long-term protective immunity. NK cells are characterized bythe expression of CD56 (both high and low levels) and the KIRfamily receptors in humans, and CD49b (DX5) and Ly-49 familymembers in mice.
NK Cell ReceptorsHuman NK Cell Receptor 58799E23359n44.1893369nneFluor 660Red LaserAPCPE-Cyanine7PerCP-eFluor 710nPEGreen, YellowGreen LasersBlue LaserFITCeFluor 450FunctionalGradeBiotinPurifiedCat. No.Clonesemi-conserved motifs (ITAM and ITIM) found in thereceptor’s intracellular domain which allows a uniqueand controlled response by the NK cell. One importantfamily of NK mediators is the activating NaturalCytotoxicity Receptors which include NKp30, NKp44and NKp46. Upon stimulation, the receptors deliverpotent signals to NK cells in order to kill target cells andproduce inflammatory cytokines such as IFNγ. NKp46 isfound on both mouse and human NK cells, while micelack a homolog for NKp44 and contain onlya pseudogene for NKp30.AntigenNK cells, unlike Natural Killer T (NKT) cells, do not expressan antigen-specific receptor. Regulation of the cytotoxicactivity of NK cells is mediated by activating and inhibitingreceptors expressed on the cell surface including naturalcytotoxicity receptors (NCR), lectin-like receptors andCD158 family of Killer Immunoglobulin-like Receptors(KIRs). These bind to specific components present on thesurface of bacteria, virally-infected cells, stressed cellsor cancer cells. Rodents lack KIRs, and instead expressfunctionally equivalent lectin-like Ly-49 receptors. Thesereceptors can be activating or inhibiting depending uponVioletLaserPerCP-Cyanine5.5Ly-49, KIR and NCR FamiliesnnnnnnnnnnnnnnnnnnnAF29-4D12 3379nnnnnnnnMouse NK Cell Receptor AntibodiesMouse NKp46Mouse NKp46 PECD56 APCCD335 PerCP-eF710Human CD335PerCP-eFluor 710Human CD56 APCMouse CD49b (DX5) APCHuman NKp30 vs NKp46Staining of normal human peripheral blood cells with Anti-Human CD56(NCAM) APC (cat. no. 17-0567) and Anti-Human CD337 (NKp30) PE(cat. no. 12-3379) (left) or Anti-Human CD335 (NKp46) PerCP-eFluor 710(cat. no. 46-3359) (right).CD335 PerCP-eF710Rat lgG2a PECD337PE PEHumanCD337Mouse CD49b (DX5) APC2Staining of C57Bl/6 splenocytes with Anti-Mouse CD49b (DX5) APC(cat. no. 17-5971) and Rat IgG2a Isotype Control PE (cat. no. 12-4321) (left)or Anti-Mouse NKp46 (29A1.4) PE (cat. no. 12-3351) (right). Total viable cellswere analyzed.Human CD56 APCCD56 9AA1 nnnnnnnnnnnLy-49C/I/F/H nnnnnnnnnnnnnnnnnnCD337 PE3
NK Cell Activation and FunctionIntracellular Flow Cytometry StainingNK cells are activated in response to IL-2, IL-12, IL-15,such as perforin and granzymes to create pores in the cellCytokine detection requires the appropriate stimulationsimultaneous analysis of surface molecules and intracellularIL-15/IL-15RA complex and IL-18, and produce and secretemembrane and initiate apoptosis via a caspase cascade inconditions and kinetics of cytokine production that will varyantigens at the single-cell level by flow cytometry. Typically, cellsa variety of cytokines, chemokines (including IFNγ, TNFα,target cells. Of the granzyme family, granzyme B is the best-depending on the cell type and the particular cytokine beingare fixed with formaldehyde to stabilize the cell membrane, andIL-17, and IL-22) and death-eliciting proteins (perforin andcharacterized, but granzymes A through M are also involvedassayed. For in vitro stimulation of cells, it is necessary to blockthen permeabilized with detergent or alcohol to create poresand have been shown to have unique ligand specificity.secretion of cytokines with protein transport inhibitors, suchin the cell membrane that require the continuous presence ofas Monensin or Brefeldin A Solution, during the final hoursthe permeabilization buffer during all subsequent steps to allowof the stimulation protocol. Simple buffer modifications allowantibodies to have access to the cytoplasm of the cell.cells contain cytoplasmic granules that contain proteinsHuman NK Intracellular Flow Cytometry AntibodiesActivate, Stimulate and Proliferate NK CellseBioscience provides high purity, low endotoxin bioactiveperformance criteria including endotoxin levels 0.01 ng/µgrecombinant proteins that can be used to activate, stimulate(tenfold lower than other suppliers) and functional testing toand proliferate NK cells. Our recombinant proteins boastsensure consistent bioactivity with every lot.the industry’s most demanding quality control andNatural Killer (NK) Differentiation ProfileCytokineHuman Cat. No. Mouse Cat. No. RoleIL-214-802934-802914-802134-8021Augments NK cell activity and boosts cytolytic activity by activating variouskinase pathwaysIL-12 p7014-812934-812914-812134-8121Induces activation, stimulates cytotoxicity and production of IFNγ and TNFIL-15/IL-15R--14-815234-8152Involved in proliferation, accumulation and survivalIL-1514-815934-815914-815334-8153Involved in proliferation, accumulation and survivalIL-18----Upregulates NK cell cytotoxicityVL19905GranulysinDH28828Granzyme BGB118899Granzyme Cy5AlexaFluor 700APC-eFluor 780eFluor 660nRed )nPerCP-eFluor 7107540Green,Yellow-GreenLasersBlue LaserFITCFL34Z3LFunctional nnnnnnnnnnnnnnnnnnnnnnnMouse NK Intracellular Flow Cytometry AntibodiesGranzyme AGzA-3G8.55831nGranzyme �XMG1.27311nLAPTW7-16B49821PerforineBioOMAK-D nnnnnnnMouse PerforinCD8 eFluor 450 CD8 eFluor 450CD8 eFluor 450mouse splenocytes were culturedmouse splenocyteswith IL-2 forwere4 daysculturedfollowedwithbyIL-25 hoursfor 4 indaysthefollowedpresencebyof5monensin.hours in theCellspresencewere thenof monensin. Cells were thenfixed and permeabilized usingfixed theandkitpermeabilized88-8824 followedusingbythestainingkit 88-8824with anti-CD8followed byandstaininganti-Perforin.with anti-CD8 and anti-Perforin.4Cat. No.Clone7539BALB/c splenocytes were stimulated for 4 days with Mouse IL-2Recombinant Protein (cat. no. 14-8021). Cells were harvested andstained with Anti-Mouse CD8a eFluor 450 (cat. no. 48-0081) followedby fixation and permeabilization using the Intracellular Fixation &Permeabilization Buffer Set and protocol (cat. no. 88-8824). Cells weresubsequently stained intracellularly with Rat IgG2a K Isotype Control APC(cat. no. 17-4321) (left) or Anti-Mouse Perforin APC (cat. no. 17-9392)(right). Cells in the lymphocyte gate were analyzed.Perforin APCT-betEOMESPFFM3Perforin APCIL-2IL-12IL-15/IL-15RIL-18CCL3(MIP-1α)Perforin APCNKPre-NKrat IgG2a APCCD56 (h)rat IgG2a APCCD49b (m)MIP-1a(CCL3)MIP-1b(CCL4)RANTES(CCL5)Granzyme BIFNγIL-17AIL-22TNFαPerforinRat IgG2a APC2R1IL-Antigenh)158(NKpNKp46CDCD11bCD161Ly49 (m)44 (h)VioletLaserAlexaFluor 488Natural Killer Cell MaturationeFluor 450granzymes). Similar to cytotoxic CD8 T cells, activated NKCD8 eFluor 450 CD8 eFluor 450CD8 eFluor 4505
NK Cell Transcriptional ControlImmunoassay QuantitationImmunoassay is a simple, effective assay platform usedto quantitatively measure either secreted or intracellularprotein biomarkers in biological samples such as serumor cell lysate. Research to gain greater insight intothese biomarker profiles will ultimately produce betterunderstandings of disease and cell biology.eBioscience produces multiple platforms for anlayteassessment and biomarker profiling—from coat-it-yourselfReady-SET-Go! ELISAs and traditional Platinum ELISAs,to ProcartaPlex Multiplex Immunoassays, using Luminex xMAP 4BP4MIP-1α (CCL3)88-7035BMS2029INSTEXP010-12029MIP-1β ANTES (CCL5)TGFβ INSTMouse ImmunoassaysGranzyme Fβ P010-20607Ready-Set-Go! ELISA – Keep costs low. Each affordable, coat-it-yourself ELISA kit includesthe reagents required to prepare and run the ELISA, including: ELISA-optimized antibodyRed LasernnnnnnnnnnnnnnnnnnnnnnnnnnnnMouse Transcription Factor AntibodiesE4BP4RANTES n, YellowGreen LaserseFluor 6Cat. lue -22ProcartaPlexSimplexPE-Cyanine7BMS2027Granzyme KELISPOTReady Set Go! PEGranzyme BInstantELISAPerCP-eFluor 710BMS2026High SensitivityELISAPerCP-Cyanine5.5Granzyme AELISAReady-Set-Go! AlexaFluor 488PlatinumELISAFITCAntigen(EOMES) and T-bet direct the fate and function ofcytotoxic cell lineages including NK cells and CD8 T cells.Furthermore, T-bet controls the developmental stability ofimmature NK cells, while EOMES regulates NK maturation.Human Transcription Factor Antibodies eFluor 450Human ImmunoassaysNK cells are the prototypical type I Innate LymphoidCells (ILC). All ILC are developmentally dependent on thetranscription factor inhibitor of DNA-binding 2 (Id2). NKcells express the transcription factor E4BP4 upstream of Id2.Additionally, the T-box transcription factors, 6502nnnnnnnnnnnnnnnnnnnnnnnnnnnnmatched pairs, standards, detection reagents, wash and coating buffers. Plates are optional.Platinum ELISA – Quantitate with confidence. Pre-coated ELISA kits provide lower inter-ProcartaPlex Multiplex Immunoassays – Quantitate more with less sample.ProcartaPlex Multiplex Immunoassays utilize Luminex xMAP technology for the biomarkerprofiling of up to 50 analytes in a single 25 μL sample. Gain a greater understanding ofC57Bl/6 splenocytes were surface stained with Anti-Mouse NK1.1 APC(cat. no. 17-5941) followed by fixation and permeabilization using theFoxp3 Buffer Set (cat. no. 00-5523) and protocol. Cells were intracellularlystained with Rat IgG2a K Isotype Control eFluor 450 (cat. no. 48-4321)(left) or Anti-Mouse EOMES eFluor 450 (cat. no. 48-4875) (right). Cells inthe lymphocyte gate were analyzed.EOMES eFluor 450EOMES eFluor 450NK1.1 APCNK1.1 APCEOMES eFluor 450The 1-wash ELISA introduces fewer handling steps, leaving more time for your research.rat IgG2a eFluor 450Instant ELISA – Reduce steps and hands-on time to increase productivity. Mouse EOMESrat IgG2a eFluor 450your research.Rat IgG2a eFluor 450and intra-assay variability with ready-to-use reagents that ensure consistent data throughoutNK1.1 APCC57 mouse splenocytes stained with NK1.1 then fixed and permed with Foxp3 buffer then IC with eomes.C57 mouse splenocytes stained with NK1.1 then fixed and permed with Foxp3 buffer then IC with eomes.NK1.1 APCNK1.1 APCNK1.1 APCbiology and disease in the same time it takes to perform an ELISA.Visit eBioscience.com/ProcartaPlex for information regarding characteristic assay detailsand a complete listing of multiplex panels and individual analytes.67
NK T CellsNK Cell Development and Maturation Transcriptional ControliNKCLP: common lymphoid progenitorCD1dCD160PLZFTCR Vα24Jα18IFNγIL-10TNFαPerforinGranzymesα 3Mags.21F70011001316019320Human EOMESneFluor 660nnnnnnnnnnnnnnnnnnnnnnnnnnnnnv alpha 24 J alpha TCR PE-Cyanine7TCRVα24Jα18v alpha24 J alphaPE-Cyanine7TCR PE-Cyanine7Mouse IgG1 PE-Cyanine7NK1.1 PE-Cy7NK1.1 PE-Cy7NK1.1 PE-Cy7CD3eFluor450 450CD3eFluorCD3eFluor450Human TCR Vα24Jα18Staining of normal human peripheral blood cells with Anti-HumanCD3 eFluor 450 (cat. no. 48-0037) and Mouse IgG1 K Isotype ControlPE-Cyanine7 (cat. no. 25-4714) (left) or Anti-Human Vα24Jα18 TCR PECyanine7 (cat. no. 25-5806) (right). Cells in the lymphocyte gate wereused for analysis.CD3eFluor450 450CD3eFluorCD3eFluor450Staining of normal human peripheral blood cells with Anti-Human CD3APC (cat. no. 17-0037) (left) or FITC (cat. no. 11-0038) (right) followed byfixation and permeabilization using the Foxp3/Transcription Factor Buffer Set(cat. no. 00-5523). Cells were then intracellularly stained with Anti-HumanEOMES FITC (cat. no. 11-4877) (left) or PerCP-eFluor 710 (cat. no. 46-4877)(right). Cells in the lymphocyte gate were analyzed.Eomes PerCP-e710EOMES PerCP-eFluor 710Eomes FITCEOMES FITCCD3 APCCD3 APCnMouse E4BP4 (NFIL3)Mouse splenocytes unstimulated (left) or stimulated overnight with MouseIL-15/IL-15R Complex Recombinant Protein Carrier-Free (cat. no. 34-8152)(right) were stained with Anti-Mouse NK1.1 PE-Cy7 (cat. no. 25-5941) andAnti-Mouse E4BP4 (NFIL3) PE (cat. no. 12-5927). Intracellular staining forE4BP4 was performed using the Foxp3 Staining Buffer Set (cat. no. 00-5523)and protocol. Cells in the lymphocyte gate were analyzed.E4BP4 PEE4BP4 PEE4BP4 PEE4BP4 PEE4BP4 PEE4BP4 PENK1.1 PE-Cy7NK1.1 PE-Cy7NK1.1 PE-Cy7L363Mouse IgG1 PE-Cyanine7MouseIgG1 PE-Cyanine7stimulated0016160993205806Red LaserMouse NKT Cell Antibodies and FormatsTranscription factors, Notch and Id2, promote commitment to type I innatelymphoid cells. Commitment to the NK cell lineage is dependent upon E4BP4and PU.1. Multiple transcription factors, Id2, T-bet and E4BP4, ensuresdevelopment from immature NK to mature NK. Finally Runx3, GATA-3, T-betand EOMES regulate mature NK cell llow-GreenLasersBlue LaserAPCAntigenImmunoregulatory(Cytokine Producing)NKCytotoxicNKFunctional GradeVioletLaserBiotinT-betEOMESHuman NKT Cell Antibodies and mNK: mature NK cell (resting)TOXPEiNK: immature NK cellPerCP-eFluor 710Id2T-betE4BP4HeliosNKP: NK cell precursorsFITCCILP: common innate lymphoid progenitorAlexaFluor 488NKPCILPCat. No.CLPIn mice, PLZF is highly expressed in immature CD1dresricted invariant NKT cells and a subset of gamma delta(Vg1.1 Vd6.3 ) T cells. In humans, PLZF is expressed in NKcells, gamma delta T cells, as well as CD4 and CD8 T cells.PLZF exists as a homodimer or in complex with PLZP, knownto be involved in the development of NKT cells, NK cellfunction, cellular quiescence and growth suppression. PLZPhas also been shown to inhibit gene expression induced byretinoic acid receptor.eFluor 450IkarosNKT cells represent a distinct lineage of T cells that expressan invariant T Cell Receptor (TCR) and share a number ofcell surface markers in common with NK cells. NKT cellsare non-polymorphic CD1d molecule-restricted T cells andare activated by glycolipid antigens presented by CD1d.Expression of CD160 and Vα24Jα18 TCR can be used toidentify mouse and human NKT cells, respectively.Expression of transcriptional repressor, PromyelocyticLeukemia Zinc Finger (PLZF), in immune cells differsbetween mice and humans.CloneNotchId2PU.1Ets-1Runx1E4BP4CD3 FITCCD3 FITC9
NK Cell Marker GuideMouse NK Cell Antibodies and 127eBioRDR51278CD161HP-3G101619CD181 (CXCR1)8F1-1-41819nCD184 (CXCR4)12F59999nnCD195 (CCR5)T21/81957nnCD197 53 (TRAIL)RIK-29927nnnCD319 (CRACC)1622229CD328 (Siglec7)QA795759CX3CR12A9-16099IL-15ReBioJM7A4 1PK1365941nnnnnNK1.1 FITCnnnnNK1.1 FITCnnnNK1.1 FITCnn2440NF-κB p65 1nnnAlexaFluor 7002B11nnnAPC-eFluor 780CD184(CXCR4)nneFluor 660CXCR3-173 3)nPECD127nPerCP eFluor r 4500621Functional GradeAntigenMEL-14CD49b(DX5)nCD244.1Normal human peripheral blood cells were left untreated (left) orstimulated for 15 min with 50 nM PMA (right). Cells were permeabilizedwith Intracellular Fixation & Permeabilization Buffer Set (cat. no. 88-8824),and then intracellularly stained with Anti-Human CD57-PE(cat. no. 12-0577-42) and Anti-Human phospho-NF-kB (Ser529)eFluor 660 (cat. no. nsstimulatedtsi tmi mu lual taet dedAlexaFluor 700APC-eFluor 780eFluor 660PE-Cyanine5.5PE-Cyanine5PerCP-eFluor 710PEnnnnneBioDM244 5837pN F k B(pS529)( S 5 2 9 ) eFluore F l u o r 660660NFkBpNFkB (S529) eFluor 660nnnC1.7p N F k B(pS529)( S 5 2 9 ) e F l u o r 660660NFkBp N F k B ( S 5 2 9 )eFluoreFluor 660PerCP-Cyanine5.5nnCD96 (TACTILE)CD57PECD57CD57PEPEnn0629unuunstimulatedsn tsi tmi mu lual taet dednnCD62L (L-Selectin) DREG56phospho NF kappaB33B4WPB p65 (S529)AlexaFluor 488FITCnnnnnRed LaserAPC0279nnnnnGreen, fiedLG.7F9nnnnnnnnnnnnnnnnnnnnnnMouse KLRG1Staining of C57Bl/6 splenocytes with Anti-Mouse NK1.1 APC(cat. no. 17-5941) and Golden Syrian Hamster IgG IsotypeControl PerCP-eFluor 710 (cat. no. 46-4914) (left) orAnti-Mouse KLRG1 PerCP-eFluor 710 (cat. no. 46-5893) (right).Total viable cells were used for analysis.KLRG1 PerCP-eFluor 710nnnBlue LasernKLRG1 PerCP-eFluor 710nnnnCat. No.0259nnn650701120161025102520253027159710491 BC96nnpolyM1/7093PC61.57D43C7LG.7F9DX5HMa2Asialo GM1CD11bCD16/32CD25KLRG1 PerCP-eFluor 710nnClonennHamster IgG PerCP-eFluor 7100168nHamster IgG PerCP-eFluor 710nnnHamster IgG PerCP-eFluor 7100113CD56eFluor 450nVioletLaserRed LaserAPCCD25Functional GradenGreen, Yellow-GreenLasersBlue RM1/5(activation epitope)CD16CB16CD27BiotinpolyPurifiedAsialo GM1Cat. No.CloneAntigenVioletLaserAlexaFluor 488Human NK Cell Antibodies and FormatsNK1.1 FITCNK1.1 FITCNK1.1 FITC11
Natural killer (NK) cells are lymphoid cells poised and ready to assist in the destruction of virally infected cells and tumor cells from the body. NK cells are part of the innate immune system that, in general, does not give rise to immunological memory or long-term protective immunity. NK cells are characterized byFile Size: 1011KB
As top predators, there is considerable research interest in the prey requirements of killer whales, so that we can evaluate their predation impact on endangered marine species and detect threats to killer whales from prey shortages. This requires information on the size, growth and body condition of killer whales.
Fly Killer Harris Home Pest Control Roach Killer Harris Home Pest Control Scorpion Killer Harris Home Pest Control Spider Killer Harris Home Pest Control Stink Bug Killer Swamp Gator Bug Barrier] {[Select Marketing Claims from "Marketing Claims" section]} Active Ingredient: Deltamethrin (CAS No. 52918-63-5) 0.03% Other .
comprised about 15 killer whales, including two adult males and at least two calves. Females and (or) subadult males pressed the attack most intently. The killer whales tore skin and blubber from the right flank of the Bryde's whale, and on 11 occasions the killer whales swam onto the head or back of the Bryde's whale, which hindered its breathing.
killer whales are seen from August to October (Reeves and Mitchell 1988a). Killer whales observed in the eastern Canadian Arctic during summer are thought to undertake seasonal migra-tions to overcome limitations imposed by pack ice (Sergeant and Fisher 1957; Reeves and Mitchell 1988a). Reeves and Mitchell (1988a) speculated killer whales
Killer Whales as Endangered and Northern Resident Killer Whales as Threatened. Both populations are listed in Schedule 1 of the Species at Risk Act (SARA). These two populations are acoustically, genetically, and culturally distinct. The "Recovery Strategy for the Northern and Southern Resident Killer Whales (Orcinus orca) in
with killer whales and other top predators (Rikardsen 2019). However, little is known about the level of overlap and the nature of interactions between killer whales and herring fishing activity in northern Nor-way. The killer whales ap pear to be attracted to fish-ing activity during the winter herring aggregations
ABSTRACT. Introduction: False killer whale (Pseudorca crassidens) is a tropical and subtropical social spe-cies that live in groups with individuals of mixed ages and sex classes. False killer whales have been documented since the late 1990s in Southwestern Costa Rica. Objective: To estimate the abundance of false killer whales in Osa Peninsula .
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