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<strong>Figure 1 Western Blot Validation in Mouse A20 Cells</strong><br>Loading: 15 μg of lysates per lane.Antibodies: IL-27 3797, (A: 1 μg/mL, B: 2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines</strong><br>Loading: 15 µg of lysates per lane.Antibodies: IL-27 3797 (4 μg/mL), IL-27 3799 (4 μg/mL), and beta-actin (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Recombinant Protein Validation </strong><br>Loading: 30 ng of human recombinant IL-17 (IL-17A+IL-17B) protein per lane.Antibodies: IL-27 3797, 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Lane 1: 0.5 μg/mL, Lane 2: 1 μg/mL, Lane 3: 2 μg/mL
<strong>Figure 4 Immunocytochemistry Validation of IL-27 in Mouse EL4 Cells</strong><br>Immunocytochemical analysis of Mouse EL4 cells using anti-IL-27 antibody (3797) at 10 μg/ml. Cells was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
Figure 1 Western Blot Validation in Mouse A20 Cells
Loading: 15 μg of lysates per lane.Antibodies: IL-27 3797, (A: 1 μg/mL, B: 2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Trial Size

IL-27 Antibody

CATALOG NUMBER: 3797

Clonality:
Polyclonal
Tested Applications:
ELISA, ICC, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Homology:
Predicted species reactivity based on immunogen sequence: Rat: (63%)
Immunogen:
Anti-IL-27 antibody (3797) was raised against a peptide corresponding to 16 amino acids near the amino terminus of human IL-27.
The immunogen is located within the first 50 amino acids of IL-27.
Conjugate:
Unconjugated
Tested Applications:
ELISA, ICC, WB
Application Note:
WB: 0.5 - 4 μg/mL; ICC: 10 μg/mL.
Antibody validated: Western Blot in human and mouse samples; Immunocytochemistry in mouse samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1288 - A20 Cell Lysate
Positive Control 2:
Cat. No. 17-207 - EL4 Cell Slide
Positive Control 3:
Cat. No. 1288 - A20 Cell Lysate
Positive Control 4:
Cat. No. 1201 - HeLa Cell Lysate
Positive Control 5:
Cat. No. 1205 - Jurkat Cell Lysate
Positive Control 6:
Cat. No. 1220 - SK-N-SH Cell Lysate
Predicted Molecular Weight:
Predicted: 28 kDa
Observed: 28 kDa
Validation:

Independent Antibody Validation in Cell lines (Figure 2) shows similar IL-27 expression profile in human and mouse cell lines detected by two independent anti-IL-27 antibodies that recognize different epitopes, 3797 against N-terminus domain and 3799 against central domain.  IL-27 proteins are detected in the most tested cell lines at different expression levels by the two independent antibodies. 

Recombinant Protein Test (Figure 3) shows that anti-IL-27 antibody (3797) detects human recombinant IL-17 (IL-17A + IL-17B) protein that contains the amino acids of 3797 antigenic peptide.

Isoforms:
Human IL-27 has one isoform (243aa, 28kD). Mouse IL-27 has one isoform (234aa, 27kD) and rat IL-27 also has one isoform (234aa, 27kD). 3797 can detect human, mouse and rat IL-27.
Purification:
IL-27 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
IL-27 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
IL-27 antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
IL27
Additional Names:
IL-27 Antibody: p28, IL30, IL-27, IL27A, IL-27A, IL27p28, Interleukin-27 subunit alpha, p28, IL-27 subunit alpha
Protein Accession Number:
NP_663634
Protein Gi Number:
28416913
Ncbi Gene Id Number:
246778
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
IL-27 Antibody: Like interleukin-23 (IL-23), IL-27 is a recently discovered member of the IL-6/IL-12 family of proinflammatory and immunoregulatory cytokines. It exists as a heterodimer composed of the p40-related protein EBI3 and an IL-12 p35-related protein termed p28. IL-27 is produced after activation by antigen-presenting cells and induces proliferation of naïve but not memory CD4+ T-cells. It acts by binding to its receptor WSX-1 and gp130 which results in the activation of a Jak/STAT signaling cascade, suggesting the IL-27 is involved in the regulation of immune processes. It has been suggested that IL-27 can also be used as a therapeutic agent against cancer as it can also induce tumor-specific anti-tumor activity mediated through CD8+ T-cells, IFN-gamma, and T-bet.
Background Reference 1:
Hunter et al. Nat. Rev. Immunol. 2005; 5:521-31.
Background Reference 2:
Pfanz et al. J. Immunol. 2004; 172:2225-31.
Background Reference 3:
Hisada et al. Cancer Res. 2004; 64:1152-6.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

3797

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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