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<strong>Figure 1 Western Blot Validation in Human, Mouse and Rat Tissues</strong><br>Loading: 15 µg of lysates per lane.Antibodies: IL-21 2465, (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP
<strong>Figure 2 Western Blot Validation with Recombinant Protein</strong><br>Loading: 30 ng of human IL-21 recombinant protein per lane.Antibodies: IL-21 2465 (0.5 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Immunohistochemistry Validation of IL-21 in Human Tonsil</strong><br> Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-IL-21 antibody (2465) at 1 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 Immunohistochemistry Validation of IL-21 in Mouse Spleen</strong><br> Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-IL-21 antibody (2465) at 1 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 5 Immunohistochemistry Validation of IL-21 in Rat Spleen</strong><br> Immunohistochemical analysis of paraffin-embedded rat spleen tissue using anti-IL-21 antibody (2465) at 1 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 6 Induced Expression Validation of IL-21 expresssion in patients with Crohn's Disease (Monteleone et al, 2007) </strong><br>Enhanced IL-21 was observed in involved but not uninvolvedCD, and was not associated with any CD phenotype, such as fibrostenosing disease. IL-21 expression was detected by anti-IL-21 antibodies (2465).
<strong>Figure 7 Regulation of IL-21 expresssion in duodenal biopsies of two patients with active Celiac disease (ACD) (Sarra et al., 2013) </strong><br>(b) shows IL-21 expression levels treated with dimethyl sulfoxide (DMSO) or wortmannin (WRT). (c) shows IL-21 expression levels with the inhibition of IL-15 antibody (aIL-15). IgG isotype was used as a control.Both WB show IL-21 expression decreased with the treatment with WRT or the inhibition of IL-15 antibody.
<strong>Figure 8 Induced Expression Validation of IL-21 expresssion in patients with Helicobacter Pylori (Hp) (Caruso et al., 2007) </strong><br>(A) shows IL-21 expression levels from biopsies of patients with Hp infection and without Hp infection (n=3).(C) shows IL-21 expression levels in CD3+LPMC cells from Hp-positive and Hp-negative patients (n=3).Both WB show IL-21 expression increased in patients with Hp infection.
Figure 1 Western Blot Validation in Human, Mouse and Rat Tissues
Loading: 15 µg of lysates per lane.Antibodies: IL-21 2465, (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP
IAV
Most Cited
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IL-21 Antibody

CATALOG NUMBER: 2465

Clonality:
Polyclonal
Tested Applications:
ELISA, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Homology:
Predicted species reactivity based on immunogen sequence: Bovine: (100%), Pig: (87%)
Immunogen:
Anti-IL-21 antibody (2465) was raised against a peptide corresponding to 15 amino acids near the center of human IL-21 precursor.
The immunogen is located within the last 50 amino acids of IL-21.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IHC-P, WB
Application Note:
WB: 0.5-2 μg/mL; IHC-P: 0.5-2 μg/mL.

Antibody validated: Western Blot and Immunohistochemistry in human, mouse and rat samples. All other applications and species not yet tested.
Specificity:
At least two isoforms of IL-21 are known to exist; this antibody will detect both isoforms. IL-21 antibody is predicted to not cross-react with other members of the interleukin protein family.
Positive Control 1:
Cat. No. 1315 – Human Tonsil Lysate
Positive Control 2:
Cat. No. 1406 – Mouse Spleen Lysate
Positive Control 3:
Cat. No. 1466 – Rat Spleen Lysate
Predicted Molecular Weight:
Predicted: 18kD
Observed: 17 kD
Validation:

Independent Antibody Validation in Cell Lines (Figure 2) shows similar IL-21 expression profile in human and mouse cell lines detected by two independent anti-IL-21 antibodies that recognize different epitopes, 2465 against central domain and 2463 against C-terminus domain.  IL-21 proteins are detected in the most tested cell lines at different expression levels by the two independent antibodies.

Recombinant Protein Test (Figure 2): Anit-IL-21 antibodies (2465) detected human IL-21 recombinant protein.

Regulated expression validation (Figure 6, 7, 8): IL-21 expression detected by anit-IL-21 antibodies (2465) was up down-regulated by WRT treatment (figure 7) or was up-regulated in Hp positive patients (figure 8) and in patients with Crohn’s disease (figure 6).

Isoforms:
Human IL-21 has 2 isoforms, including isoform 1 (162aa, 18.7kD) and isoform 2 (153aa, 17.6kD). Mouse IL-21 has one isoform (146aa, 16.8kD) and Rat IL-21 also has one isoform (146aa, 17.1kD). 2465 can detect both human isoforms and also detect mouse and rat.
Purification:
IL-21 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
IL-21 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/ml
Storage Conditions:
IL-21 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:
IL21
Additional Names:
IL-21 Antibody: Za11, IL-21, Interleukin-21, Za11
Protein Accession Number:
NP_068575
Protein Gi Number:
11141875
Ncbi Gene Id Number:
59067
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
IL-21 Antibody: A novel cytokine was recently identified in human and mouse and designated IL-21, which has significant homology to IL-2, IL-4, and IL-15. The receptor for IL-21 (IL-21R, also termed NILR for novel Interleukin receptor) is a new member of the class I cytokine receptor family. IL-21R forms a complex with the common cytokine receptor gamma chain, gammac, and mediates IL-21 signaling. IL-21 and its receptor activate JAK-STAT signaling pathway. IL-21 is expressed in activated T cells, and HL-60 and THP-1 cell lines. IL-21 plays a role in the proliferation and maturation of NK, B and T cell populations.
Background Reference 1:
Parrish-Novak et al. Nature 2000; 408:57-63.
Background Reference 2:
Ozaki et al. Proc. Natl. Acad. Sci. USA. 2000; 97:11439-44.
Background Reference 3:
Asao et al. J. Immunol. 2001; 167:1-5.
Citation 1:
Monteleone et al. Interleukin-21 enhances T-helper cell type I signaling and interferon-γ production in Crohn's disease. Gastroenterology. 2005;128(3):687-94. PMID: 15765404
Citation 2:
Sarra et al. IL-15 positively regulates IL-21 production in celiac disease mucosa. Mucosal Immunol. 2013;6(2):244-55. PMID: 22785229
Citation 3:
Caruso et al. IL-21 is highly produced in Helicobacter pylori-infected gastric mucosa and promotes gelatinases synthesis. J Immunol. 2007;178(9):5957-65. PMID: 17442980
Citation 4:
Daniele Fina. Interleukin-21 controls inflammatory pathways in IBD. University of Rome, Italy. PhD thesis. 2009.PMID:
Citation 5:
Roberta Caruso. Expression and functional role of Th-17 cells in helicobacter pylori-related gastritis. University of Rome, Italy. PhD thesis. 2008.PMID:
Citation 6:
Giovanni Monteleone. Antigenicepitopes of Interleukin-21, related antibodies and their use in medical field. U.S patent application publication. 2009.PMID:

FOR RESEARCH USE ONLY

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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:

2465

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

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