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<strong>Figure 1 Western Blot Validation in Rat Spleen Tissue</strong><br>Loading: 15 μg of lysates per lane.Antibodies: EBI-3 4977 (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 Recombinant Protein Validation </strong><br>Loading: 30 ng of human recombinant IL-17 (IL-17A+IL-17B) protein per lane.Antibodies: EBI-3 4977, 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 3 Immunohistochemistry Validation of EBI-3 in Human Spleen Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded Human Spleen tissue using anti-EBI-3 antibody (4977) at 2.5 μg/ml. Tissue 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.
<strong>Figure 4 Immunofluorescence Validation of EBI-3 in Human Spleen Tissue</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed Human Spleen Tissue labeling EBI-3 with 4977 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
Figure 1 Western Blot Validation in Rat Spleen Tissue
Loading: 15 μg of lysates per lane.Antibodies: EBI-3 4977 (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Trial Size

EBI3 Antibody

CATALOG NUMBER: 4977

Clonality:
Polyclonal
Tested Applications:
ELISA, IF, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Immunogen:
Anti-EBI-3 antibody (4977) was raised against a peptide corresponding to 16 amino acids near the center of mature human EBI-3.
The immunogen is located within amino acids 150 - 200 of EBI3.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IF, IHC-P, WB
Application Note:
WB: 0.5 - 2 μg/mL; IF: 20 μg/mL ; IHC: 2.5 μg/mL;.
Antibody validated: Western Blot in human and rat samples; Immunofluorescence and Immunohistochemistry in human samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1466 - Rat Spleen Tissue Lysate
Positive Control 2:
Cat. No. 10-901 - Human Spleen Tissue Slide
Predicted Molecular Weight:
Predicted: 25kD
Observed: 30 kD (25kD + 2 N-Glycosylations)
Validation:

Recombinant Protein Test (Figure 2) shows that anti-IL-EBI-3 antibody (4977) detects human recombinant IL-17 (IL-17A + IL-17B) protein that contains the amino acids of 4977 antigenic peptide.

 

Isoforms:
Human EBI-3 has only 1 isoform (229aa, 25kD). Mouse EBI-3 has one isoform (228aa, 25kD) and rat EBI-3 also has one isoform (228aa, 25kD). 4977 can detect human isoform and possibly detect mouse and rat EBI-3.
Purification:
EBI3 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
EBI3 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
EBI3 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:
EBI3
Additional Names:
EBI3 Antibody: IL27B, IL-27B, IL27B, Interleukin-27 subunit beta, Epstein-Barr virus-induced gene 3 protein, IL-27 subunit beta
Protein Accession Number:
NP_005746
Protein Gi Number:
14577917
Ncbi Gene Id Number:
10148
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
EBI3 Antibody: EBI3 is a subunit in two distinct heterodimeric cytokines: interleukin-27 (IL-27) and IL-35. 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 (also known as TCCR) 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. IL-35 is composed of EBI3 and the p35 subunit of IL-12 and has been reported to have therapeutic effects against collagen-induced arthritis by expanding the population of regulatory T cells and suppressing Th17 cells. At least two isoform of EBI3 are known to exist.
Background Reference 1:
Pfanz et al. Immunity2002; 16:779-90.
Background Reference 2:
Devergne et al. Proc. Natl. Acad. Sci. USA1997; 94:12041-6.
Background Reference 3:
Niedbala  et al. Eur. J. Immunol.2007; 3021-29.
Background Reference 4:
Pfanz et al. J. Immunol.2004; 172:2225-31.

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:

4977

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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