Immunofluorescence analysis of IL-8 was performed using 70% confluent log phase U-87 MG cells treated 1 uM thapsigargin for 24 hours. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with IL-8 (3IL8-H10) Mouse Monoclonal Antibody (M801) at 2µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjµgate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e is untreated cell with no signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification..
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG|
|Immunogen||Recombinant human IL-8|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
M801 targets IL-8 in ELISA, and WB applications and shows reactivity with Human samples.
The M801 immunogen is recombinant human IL-8.
M801 detects IL-8 which has a predicted molecular weight of approximately 9 kDa.
The M801 IL8 antibody (clone 3IL8-H10) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M802B (biotinylated conjugate of clone I8-S2). Typical dilutions for sandwich ELISA include 1 µg/ml for coating and 0.125 - 0.25 µg/ml for detection.
Antibody M801 (clone 3IL8-H10) and biotinylated antibody M802B (clone I8-S2) have successfully been used in combination with recombinant IL8 protein SIL8 in ELISA applications.
The protein encoded by this gene is a member of the CXC chemokine family. This chemokine is one of the major mediators of the inflammatory response. This chemokine is secreted by several cell types. It functions as a chemoattractant, and is also a potent angiogenic factor. This gene is believed to play a role in the pathogenesis of bronchiolitis, a common respiratory tract disease caused by viral infection. This gene and other ten members of the CXC chemokine gene family form a chemokine gene cluster in a region mapped to chromosome 4q.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Transforming growth factor-ß impairs glucocorticoid activity in the A549 lung adenocarcinoma cell line.
M801 was used in ELISA to investigate the effect of transforming growth factor-beta on glucocorticoid activity in human A549 cells
|Salem S,Harris T,Mok JS,Li MY,Keenan CR,Schuliga MJ,Stewart AG||British journal of pharmacology (166:2036)||2012|
Ocular fibroblast diversity: implications for inflammation and ocular wound healing.
M801 was used in ELISA to study the significance of ocular fibroblast diversity for inflammation and ocular wound healing
|Xi X,McMillan DH,Lehmann GM,Sime PJ,Libby RT,Huxlin KR,Feldon SE,Phipps RP||Investigative ophthalmology and visual science (52:4859)||2011|
Inhibitors of p38 suppress cytokine production in rheumatoid arthritis synovial membranes: does variable inhibition of interleukin-6 production limit effectiveness in vivo?
M801 was used in ELISA to investigate the role of p38 kinase in rheumatoid arthritis synovial membranes
|Page TH,Brown A,Timms EM,Foxwell BM,Ray KP||Arthritis and rheumatism (62:3221)||2010|
Intermittent haemodiafiltration in refractory congestive heart failure: BNP and balance of inflammatory cytokines.
M801 was used in ELISA to assess the interleukin-8 level in patients with decompensated congestive heart failure
|Libetta C,Sepe V,Zucchi M,Pisacco P,Cosmai L,Meloni F,Campana C,Rampino T,Monti C,Tavazzi L,Dal Canton A||Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association (22:2013)||2007|
IgE modulates neutrophil survival in asthma: role of mitochondrial pathway.
M801 was used in ELISA to study the role of mitochondrial pathway in neutrophil survival
|Saffar AS,Alphonse MP,Shan L,Hayglass KT,Simons FE,Gounni AS||Journal of immunology (Baltimore, Md. : 1950) (178:2535)||2007|
Protein microarray platform for the multiplex analysis of biomarkers in human sera.
M801 was used in ELISA to develop a protein microarray platform for the multiplex analysis of biomarkers in human sera
|Urbanowska T,Mangialaio S,Zickler C,Cheevapruk S,Hasler P,Regenass S,Legay F||Journal of immunological methods (316:1)||2006|
|Human||1 ug ml||
Oxidized ATP (oATP) attenuates proinflammatory signaling via P2 receptor-independent mechanisms.
M801 was used in ELISA to study the role of oxidized ATP in proinflammatory signaling
|Beigi RD,Kertesy SB,Aquilina G,Dubyak GR||British journal of pharmacology (140:507)||2003|
Development of protein microarray technology to monitor biomarkers of rheumatoid arthritis disease.
M801 was used in ELISA to develop protein microarray technology to monitor biomarkers of rheumatoid arthritis disease
|Urbanowska T,Mangialaio S,Hartmann C,Legay F||Cell biology and toxicology (19:189)||2003|
Regulation of interleukin 8 expression in human malignant melanoma cells.
M801 was used in ELISA to investigate the regulation of interleukin 8 expression in melanoma cells
|Singh RK,Varney ML||Cancer research (58:1532)||1998|
Antineutrophil cytoplasmic antibodies induce monocyte IL-8 release. Role of surface proteinase-3, alpha1-antitrypsin, and Fcgamma receptors.
M801 was used in ELISA to investigate the role of surface proteinase-3, alpha1-antitrypsin, and Fcgamma receptors in monocyte interleukin 8 release
|Ralston DR,Marsh CB,Lowe MP,Wewers MD||The Journal of clinical investigation (100:1416)||1997|
Secretion of proinflammatory cytokines by epithelial cells in response to Chlamydia infection suggests a central role for epithelial cells in chlamydial pathogenesis.
M801 was used in ELISA to investigate the important roles of epithelial cells in inflammatory response to chlamydia
|Rasmussen SJ,Eckmann L,Quayle AJ,Shen L,Zhang YX,Anderson DJ,Fierer J,Stephens RS,Kagnoff MF||The Journal of clinical investigation (99:77)||1997|
Monocyte Fc gamma receptor cross-linking induces IL-8 production.
M801 was used in ELISA to investigate the influence of monocyte Fc gamma receptor cross-linking on the expression of interleukin 8
|Marsh CB,Gadek JE,Kindt GC,Moore SA,Wewers MD||Journal of immunology (Baltimore, Md. : 1950) (155:3161)||1995|
Rapamycin treatment depresses intragraft expression of KC/MIP-2, granzyme B, and IFN-gamma in rat recipients of cardiac allografts.
M801 was used in immunohistochemistry to investigate the effects of rapamycin on KC/MIP-2, granzyme B, and interferon gamma expression on cardiac allografts
|Wieder KJ,Hancock WW,Schmidbauer G,Corpier CL,Wieder I,Kobzik L,Strom TB,Kupiec-Weglinski JW||Journal of immunology (Baltimore, Md. : 1950) (151:1158)||1993|