Anti-IL-8 抗体

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-8 antibody (AB106350)

IL8 was detected in a paraffin-embedded section of human stomach cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml ab106350 overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using a HRP conjugated Rabbit IgG with DAB as the chromogen.

• WB

Supplier Data

Western blot - Anti-IL-8 antibody (AB106350)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, ab322732 and ab106350 were shown to bind specifically to IL-8. Target of interest was observed at 11 kDa in untreated wild-type PC-3 cell lysates (lane 1) and upregulated in treated wild-type PC-3 cell lysates (lane 2). No signal was observed at this size in untreated IL-8 knockout cell line (lane 3) or treated IL-8 knockout cell line (lane 4).

Recombinant multiclonal ab322732 showed higher sensitivity than polyclonal ab106350.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-IL-8 antibody [RM1267] (<a href='/products/primary-antibodies/il-8-antibody-rm1267-ab322732'>ab322732</a>) at 1/1000 dilution

Lane 1:

Untreated Wild-type PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Wild-type PC-3 treated with2 ug/ml LPS for 1 hour, 5ug/ml Brefeldin A was then added for additional 5 hours, whole cell lysate at 20 µg

Lane 3:

Untreated CXCL8 knockout PC-3 whole cell lysate at 20 µg

Lane 4:

CXCL8 knockout PC-3 treated with 2 ug/ml LPS for 1 hour, 5ug/ml Brefeldin A was then added for additional 5 hours, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 11 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-IL-8 antibody (AB106350)

All lanes:

Western blot - Anti-IL-8 antibody (ab106350) at 0.5 µg/mL

All lanes:

Recombinant human IL8 protein at 10 ng

Secondary

All lanes:

Goat anti-rabbit IgG-HRP at 1/5000 dilution

false

• WB

CiteAb

Western blot - Anti-IL-8 antibody (AB106350)

Western Blotting using Anti-IL-8 antibody, ab106350. Publication image from Allouch, A. et al., 2022, Nat Commun, 36347876. Legend direct from paper.

Tumor cell phagocytosis triggers the proinflammatory activation of macrophages.a Schematic representation of the coculture of CMFDA-labeled MDMs with CMTMR-labeled malignant hematologic cells. b Confocal micrograph of MDMs and Jurkat cells after 8 h of coculture. c, d Percentage of phagocytosis of leukemia cells or PBLs (c) or CD34+ AML cells (d) in control (Co.)- or ZVAD (100 µM)-treated cocultures. e FACS dot plot of Phago+ MDMs or Phago- MDMs sorted after 2 h of coculture with MOLT4 cells. f, g Confocal micrographs and percentages of CMTMR+CMFDA+ MDMs at 2 h (**p = 0.0079) (f, g) and 96 h (h, i) after Phago+ MDMs and Phago- MDMs sorting. j–n Phago+ MDMs were analyzed in comparison to Phago- MDMs to characterize modulated genes by a microarray (**p = 0.0022, *p = 0.0152) (j), CD163 membrane expression by FACS (**p = 0.0039) (k), and IRF5 expression by western blot (WB) analysis (l) and the supernatant (SN) was evaluated for indicated proinflammatory cytokines by WB analysis (l) or for IFNγ by a cytokine microarray (*p = 0.0286) (m) or ELISA (***p = 0.0008) (n) at 96 h (j, k, m) and 7 d (l, n) after FACS sorting. o, p Transwell coculture model of Phago+ MDMs and Phago- MDMs at 2 h after FACS sorting (o), and iNOS expression identified by WB analysis of Phago- MDMs cocultured in the bottom chambers for 15 d (p). In b, l, p and e, f, h, the data are representative of n = 3 and n = 5 donors. In c, n, d, and g, i, the data are presented as the mean±SEM from n = 3, n = 6, and n = 5 donors. In d, the CD34+ cells were from n = 4 AML patients. In j, box plots show centre line as median, box limits as upper and lower quartiles, and whiskers as a minimum to maximum values, from n = 3 donors. In k and m, the data are donor matched from n = 9 and n = 4 donors. Exact p-values are indicated and determined with two- (g) or one-tailed (m) unpaired Mann-Whitney test, the Kolmogorov–Smirnov test (j), a two-tailed paired Wilcoxon test (k), and a two-tailed unpaired t test (n). Source data are provided as a Source Data file.

false