Revision 6

#94407Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-P, IF-IC, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

11

Source/Isotype:

Rabbit IgG

UniProt ID:

#P10145

Entrez-Gene Id:

3576

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunohistochemistry (Paraffin) 1:50 - 1:200
Immunofluorescence (Immunocytochemistry) 1:200 - 1:800
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier-free (BSA and azide free) version of this product see product #10681.

Specificity / Sensitivity

IL-8 (E5F5Q) XP® Rabbit mAb recognizes endogenous levels of total IL-8 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with human IL-8 recombinant protein.

Background

The prototypical CXC chemokine, IL-8, is best known for effects on neutrophils, specifically its ability to act as a chemoattractant and activate degranulation and respiratory burst (1-3).

IL-8 is produced by a number of cell types, including monocytes, T cells, neutrophils, fibroblasts, endothelial cells, and others (1). In addition to its effects on neutrophils, IL-8 promotes angiogenesis, inhibits endothelial cell apoptosis, and promotes the proliferation of melanoma cells in an autocrine fashion (1). As a result, IL-8 may be a contributing factor to the development of certain cancers (1,2). There are two distinct receptors for IL-8, CXCR1 and CXCR2; both are G protein-coupled receptors (1). Ligand binding induces Ca2+ mobilization and activates the PI3K, PKC, Rho, and Rac pathways (1,3).

  1. Payne, A.S. and Cornelius, L.A. (2002) J Invest Dermatol 118, 915-22.
  2. Brat, D.J. et al. (2005) Neuro Oncol 7, 122-33.
  3. Mukaida, N. (2003) Am J Physiol Lung Cell Mol Physiol 284, L566-77.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 6
#94407

IL-8 (E5F5Q) XP® Rabbit mAb

Western Blotting Image 1: IL-8 (E5F5Q) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from U-937 cells, untreated (-) or treated (+) as indicated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (100 ng/mL, 24 hr), Lipopolysaccharides (LPS) #14011 (5 μg/ml, 7 hr), and/or Brefeldin A #9972 (300 ng/mL, 3 hr), using IL-8 (E5F5Q) XP® Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 2: IL-8 (E5F5Q) XP® Rabbit mAb Expand Image
Western blot analysis of recombinant Human Interleukin-8 (hIL-8) #94853 using IL-8 (E5F5Q) XP® Rabbit mAb.
Immunoprecipitation Image 1: IL-8 (E5F5Q) XP® Rabbit mAb Expand Image
Immunoprecipitation of IL-8 protein from extracts of U-937 cells treated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (100 ng/mL, 24 hr), Lipopolysaccharides (LPS) #14011 (5 μg/ml, 7 hr), and Brefeldin A #9972 (300 ng/mL, 3 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is IL-8 (E5F5Q) XP® Rabbit mAb. Western blot analysis was performed using IL-8 (E5F5Q) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
Immunohistochemistry Image 1: IL-8 (E5F5Q) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using IL-8 (E5F5Q) XP® Rabbit mAb.
Immunohistochemistry Image 2: IL-8 (E5F5Q) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma (left) or endometrioid adenocarcinoma (right) using IL-8 (E5F5Q) XP® Rabbit mAb (top) or IL-8 Rabbit mAb (bottom). These two antibodies detect unique, non-overlapping epitopes on human IL-8. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Immunohistochemistry Image 3: IL-8 (E5F5Q) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human esophageal carcinoma using IL-8 (E5F5Q) XP® Rabbit mAb.
Immunohistochemistry Image 4: IL-8 (E5F5Q) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human tonsil using IL-8 (E5F5Q) XP® Rabbit mAb.
Immunohistochemistry Image 5: IL-8 (E5F5Q) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using IL-8 (E5F5Q) XP® Rabbit mAb.
Immunohistochemistry Image 6: IL-8 (E5F5Q) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human small cell lung carcinoma using IL-8 (E5F5Q) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right). 
Immunohistochemistry Image 7: IL-8 (E5F5Q) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded U-937 cell pellet, untreated (left, negative) or treated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (100 ng/mL, 24 hr), Lipopolysaccharides (LPS) #14011 (5 μg/ml, 7 hr), and Brefeldin A #9972 (300 ng/mL, 3 hr) (right, positive), using IL-8 (E5F5Q) XP® Rabbit mAb.
Immunohistochemistry Image 8: IL-8 (E5F5Q) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded 293T cell pellet, untransfected (left) or IL8-transfected (right), using IL-8 (E5F5Q) XP® Rabbit mAb.
Immunofluorescence Image 1: IL-8 (E5F5Q) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of U-937 cells, untreated (left, negative) or treated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (100 ng/mL, 24 hr) and Lipopolysaccharides (LPS) #14011 (5 μg/ml, 7 hr; right, positive), using IL-8 (E5F5Q) XP® Rabbit mAb (green) and DAPI #4083 (blue).
Flow Cytometry Image 1: IL-8 (E5F5Q) XP® Rabbit mAb Expand Image
Flow cytometric analysis of U-937 cells, untreated (blue) or treated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (100 ng/mL, 24 hr), Lipopolysaccharides (LPS) #14011 (5 μg/ml, 7 hr), and Brefeldin A #9972 (300 ng/mL, 3 hr) (green), using IL-8 (E5F5Q) XP® Rabbit mAb (solid lines) or concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.