MA5-23697

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)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-23697

Provider

Invitrogen Antibodies

Product name

IL-8 (CXCL8) Monoclonal Antibody (6217)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

In Western blots, this antibody shows 100% cross-reactivity with recombinant porcine CXCL8/IL-8 and no cross-reactivity with rrCXCL3/CINC-2 alpha.

Antibody clone number

6217

Concentration

0.5 mg/mL

References

Submitted references

Tumour cell apoptosis modulates the colorectal cancer immune microenvironment via interleukin-8-dependent neutrophil recruitment.

Schimek V, Strasser K, Beer A, Göber S, Walterskirchen N, Brostjan C, Müller C, Bachleitner-Hofmann T, Bergmann M, Dolznig H, Oehler R
Cell death & disease 2022 Feb 4;13(2):113

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FABP4 deactivates NF-κB-IL1α pathway by ubiquitinating ATPB in tumor-associated macrophages and promotes neuroblastoma progression.

Miao L, Zhuo Z, Tang J, Huang X, Liu J, Wang HY, Xia H, He J
Clinical and translational medicine 2021 Apr;11(4):e395

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-IL-8 (CXCL8) Monoclonal Antibody (6217)(Product # MA5-23697) and a 11kDa band corresponding to IL-8 (CXCL8) was observed in U-87 MG up on treatment with Thapsigargin and PTI. Whole cell extracts (30 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 Dura Extended Duration Substrate (Product # 34076).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of IL-8 (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 (CXCL8) Monoclonal Antibody (6217) (Product # MA5-23697) at 8 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.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemical analysis of CXCL8/IL-8 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using human CXCL8/IL-8 Monoclonal Antibody (Product # MA5-23697) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the 557-conjugated Anti-mouse IgG Secondary Antibody (re and counterstained with DAPI (blue).

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 4 Neutrophils co-localise with apoptotic tumour cells and tumour-associated macrophages in CRC pseudolumina. A Flow cytometric analysis of dissociated CRC tissues and matched normal mucosae of treatment-naive CRC patients. For gating strategy, see Fig. S7A . Graphs present median and 95% CI (%CD66b+ cells, n = 15) or mean +- SD (surface marker MFIs, n = 10-12). * P < 0.05, ** P < 0.01, ns not significant, as calculated by Wilcoxon signed-rank test (%CD66b+ cells) or two-tailed paired t -tests (surface marker MFIs). B Representative CD66b immunohistochemistry (IHC) of a CRC tissue. Red and black arrows indicate stromal and intraluminal neutrophils, respectively, while dotted lines illustrate CRC pseudolumina. Scale bars, 100 um. C , D Consecutive IHC sections of representative CRC specimens stained for M30, cleaved caspase-3 and CD66b ( C upper panel), CD3, CD68 and CD66b ( C lower panel) and IL-8 and CD66b ( D ). Arrows indicate positive staining in CRC pseudolumina. Scale bars, 100 um. E Quantification of cleaved caspase-3-expressing, M30-expressing, IL-8-expressing, CD66b-expressing, CD68-expressing and CD3-expressing CRC pseudolumina in IHC sections of 35 CRC patients. Patients with at least 20 IHC-positive pseudolumina/cm 2 tissue area were considered positive in the presented Venn diagrams. F CD66b and M30 immunofluorescence staining of a CRC pseudolumen. Arrows indicate close contact between CD66b-positive neutrophils and M30-positive apoptotic tumour cells. Images