Sandwich ELISA analysis of human IL-8 was performed using a Human IL-8 Colorimetric ELISA kit (Product # EH2IL8) by loading 50 ul per well of Human IL-8 Recombinant Protein (Product # SIL8) in dodecuplicate at 1000, 400, 160, 64, 25.6, and 0 pg/ml across a 3 ug/ml mouse anti-Human IL-8 (Product # M801) pre-coated plate and incubating for 1 hour at room temperature. The plate was washed, and then incubated with 50 ul per well of a biotinylated mouse anti-human IL-8 monoclonal antibody (Product # M802B) in duplicate at 0.25 ug/ml for 1 hour at room temperature. The plate was washed and incubated with 100 ul per well of Streptavidin-HRP (Product # N504) in all test wells at a 1:4,000 dilution for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 30 minutes at room temperature in the dark. The plate was then stopped with 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG2b, kappa|
|Immunogen||Recombinant human IL-8|
|Storage buffer||PBS with 4% BSA|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|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.
|ELISA (ELISA)||See 7 publications below|
M802B targets IL-8 in ELISA, and WB applications and shows reactivity with Human samples.
The M802B immunogen is recombinant human IL-8.
M802B detects IL-8 which has a predicted molecular weight of approximately 9 kDa.
The M802B IL8 antibody (biotinylated conjugate of clone I8-S2) has successfully been paired as the detection antibody in a sandwich ELISA with capture antibody M801 (clone 3IL8-H10). Typical dilutions for sandwich ELISA include 1 µg/ml for coating and 0.125 - 0.25 µg/ml for detection.
Biotinylated antibody M802B (clone I8-S2) and antibody M801 (clone 3IL8-H10) 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.
M802B 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|
Optical protein sensor for detecting cancer markers in saliva.
M802B was used in ELISA to develop a new sensor for cancer detection in saliva
|Tan W,Sabet L,Li Y,Yu T,Klokkevold PR,Wong DT,Ho CM||Biosensors and bioelectronics (24:266)||2008|
IgE modulates neutrophil survival in asthma: role of mitochondrial pathway.
M802B 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.
M802B 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|
Oxidized ATP (oATP) attenuates proinflammatory signaling via P2 receptor-independent mechanisms.
M802B 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.
M802B 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.
M802B was used in ELISA to investigate the regulation of interleukin 8 expression in melanoma cells
|Singh RK,Varney ML||Cancer research (58:1532)||1998|