IL-21 Antibody

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ProSci
Product Code: PSI-2465
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-2465-0.02mg0.02mg£150.00
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PSI-2465-0.1mg0.1mg£449.00
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Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunohistochemistry (IHC)
  • Western Blot (WB)

Images

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<strong>Figure 1 Western Blot Validation in Human, Mouse and Rat Cell Lines</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 conjugate at 1:10000 dilution.
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<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-21 2465 (1 μg/mL), IL-21 2463 (5 μg/mL), beta-actin (5 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.
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<strong>Figure 3 Western Blot Validation in Human HL-60 Cell Lysate</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 conjugate at 1:10000 dilution. (A) Absence of blocking peptide (B) Presence of blocking peptide.
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<strong>Figure 4 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.
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<strong>Figure 5 Immunocytochemistry Validation of IL-21 in K562 Cells</strong><br> Immunocytochemical analysis of K562 cells using anti-IL-21 antibody (2465) at 2 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.
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<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 uninvolved CD, and was not associated with any CD phenotype, such as fibrostenosing disease. IL-21 expression was detected by anti-IL-21 antibodies (2465).
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<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.
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<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.
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<strong>Figure 1 Western Blot Validation in Human, Mouse and Rat Cell Lines</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 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-21 2465 (1 μg/mL), IL-21 2463 (5 μg/mL), beta-actin (5 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 Western Blot Validation in Human HL-60 Cell Lysate</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 conjugate at 1:10000 dilution. (A) Absence of blocking peptide (B) Presence of blocking peptide.
<strong>Figure 4 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 5 Immunocytochemistry Validation of IL-21 in K562 Cells</strong><br> Immunocytochemical analysis of K562 cells using anti-IL-21 antibody (2465) at 2 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.
<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 uninvolved CD, 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.

Documents

Further Information

Additional Names:
IL-21 Antibody: Za11, IL-21, Interleukin-21, Za11
Application Note:
WB: 1 μg/mL; ICC: 2 μg/mL.

Antibody validated: Western Blot in human, mouse and rat samples; Immunocytochemistry in human samples. All other applications and species not yet tested.
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 References:
  • Parrish-Novak et al. Nature 2000; 408:57-63.
  • Ozaki et al. Proc. Natl. Acad. Sci. USA. 2000; 97:11439-44.
  • Asao et al. J. Immunol. 2001; 167:1-5.
Buffer:
IL-21 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/ml
Conjugate:
Unconjugated
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.
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.
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.
NCBI Gene ID #:
59067
NCBI Official Name:
interleukin 21
NCBI Official Symbol:
IL21
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 18kD

Observed: 17 kD
Protein Accession #:
NP_068575
Protein GI Number:
11141875
Purification:
IL-21 Antibody is affinity chromatography purified via peptide column.
Research Area:
Chemokines & Cytokines,Cancer
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.
Swissprot #:
Q9HBE4
User NOte:
Optimal dilutions for each application to be determined by the researcher.
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 4): 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).

References

  1. Monteleone et al. Interleukin-21 enhances T-helper cell type I signaling and interferon-? production in Crohn?s disease. Gastroenterology. 2005;128(3):687-94. PMID: 15765404
  2. Sarra et al. IL-15 positively regulates IL-21 production in celiac disease mucosa. Mucosal Immunol. 2013;6(2):244-55. PMID: 22785229
  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
  4. Daniele Fina. Interleukin-21 controls inflammatory pathways in IBD. University of Rome, Italy. PhD thesis. 2009.PMID:
  5. Roberta Caruso. Expression and functional role of Th-17 cells in helicobacter pylori-related gastritis. University of Rome, Italy. PhD thesis. 2008.PMID:
  6. Giovanni Monteleone. Antigenicepitopes of Interleukin-21, related antibodies and their use in medical field. U.S patent application publication. 2009.PMID:

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