14-7189-82

antibody from Invitrogen Antibodies

Targeting: CXCL8 3-10C, AMCF-I, b-ENAP, GCP-1, GCP1, IL-8, IL8, K60, LECT, LUCT, LYNAP, MDNCF, MONAP, NAF, NAP-1, NAP1, SCYB8, TSG-1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

14-7189-82

Provider

Invitrogen Antibodies

Product name

IL-8 (1-77) (CXCL8) Monoclonal Antibody (N11), eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The monoclonal antibody N11 reacts specifically with the 77 amino acid form of IL-8 (IL-877, IL-8(1-77), CXCL877) and does not react with the more common 72 amino acid form. Frequently referred to as "endothelial" IL-8, the 77 amino acid form is secreted in cultured leukocytes. The 72 amino acid form is the result of efficient cleavage of an N-terminal pentamer from IL-877 after secretion. IL-8 is a C-X-C chemokine with potency in neutrophil chemotaxis and activation. IL-877 has decreased chemotactic activity when compared to IL-872, but has other specific pro-apoptotic and tumor cell chemotactic activities. IL-877 has recently been shown to be the major isoform present in premature neonates due to deficiency in serum protease activity that is not seen in term neonates.

Antibody clone number

N11

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoblot of human peripheral blood mononuclear cells either left untreated or treated with LPS and Brefeldin A for 4 hours. Lane 1, non-reduced lysate prepared from stimulated cells; lane 2, non-reduced lysate of unstimulated cells; lane 3, reduced lysate of stimulated cells; lane 4, reduced lysate prepared from unstimulated cells. Bands were visualized using Anti-Mouse IgG HRP.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-IL-8 (1-77) (CXCL8) Monoclonal Antibody (N11), eBioscience™(Product # 14-7189-82) and a 11kDa band corresponding to IL-8 (1-77) (CXCL8) was observed in U-87 MG up on treatment with Thapsigargin and PTI. Whole cell extracts (60 µg lysate) of U-87 MG (Lane 1), U-87 MG treated with PTI (1X for 4hr) (Lane 2), U-87 MG treated with Thapsigargin (1uM for 24hr) followed by PTI (1X for 4hr) (Lane 3) were electrophoresed using Novex™ 16% Tricine Protein Gel (Product # EC6695BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of IL-8 (1-77) (CXCL8) was performed using 70% confluent log phase U-87 MG cells treated with 1uM of Thapsigargin for 24 hours. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with IL-8 (1-77) (CXCL8) Monoclonal Antibody (N11), eBioscience™ (Product # 14-7189-82) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic(Golgi complex like pattern) localization. Panel e represents untreated cells . Panel f represents cells treated with PTI . Panel g represents cells treated with Thapsigargin and PTI. Panel h represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.