ERN1 Antibody

• Catalog Number: orb1239523
• Category: Antibodies
• Description: ERN1 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, ICC, IF, IHC-P, WB
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: Anti-IRE1p antibody (orb1239523) was raised against a peptide corresponding to 16 amino acids near the carboxy terminus of human IRE1P. The immunogen is located within the last 50 amino acids of IRE1p.
• Concentration: 1 mg/mL
• Dilution range: WB: 0.5-2 μg/mL; ICC: 1 μg/mL; IF: 2-20 μg/mL; IHC: 2 μg/mL.Antibody validated: Western Blot in human, mouse and rat samples; Immunocytochemistry in mouse samples; Immunofluorescence in human, mouse and rat samples; Immunohistochemistry in human and rat samples. All other applications and species not yet tested.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: Predicted: 110kDObserved: 110kD
• Target: ERN1
• UniProt ID: O75460
• NCBI: O75460
• Storage: IRE1p 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.
• Buffer/Preservatives: IRE1p Antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: IRE1p Antibody: IRE1, IRE1P, IRE1a, hIRE1p, IRE1,
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Western Blot Validation in Mouse A20 Cell Lysate. Loading: 15 µg of lysates per lane. Antibodies: IRE1p orb1239523 (A: 0.5 µg/mL, B: 1 µg/mL, C: 2 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

KO Validation in HeLa Cells. Loading: 10 µg of WT cell lysates (lane 1) or IRE1P KO cell lysates (lane 2). Antibodies: IRE1P orb1239523 (0.5 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.

Western Blot Validation in Human Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: IRE1p orb1239523 (0.4 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: Caco-2, Lane2: SK-N-SH.

Western Blot Validation in Rat Brain Tissue Lysate. Loading: 15 µg of lysates per lane. Antibodies: IRE1p orb1239523 (A: 0.5 µg/mL, B: 1 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Immunofluorescence Validation of IRE1p in Mouse A20 Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed A20 Cells labeling IRE1P with orb1239523 at 2 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).

Immunocytochemistry Validation of IRE1p in Mouse A20 Cells. Immunocytochemical analysis of A20 cells using anti-IRE1p antibody (orb1239523) at 1 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.

Immunofluorescence Validation of IRE1p in Human Small Intestine Tissue. Immunofluorescent analysis of 4% paraformaldehyde-fixed Human Small Intestine Tissue labeling IRE1p with orb1239523 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Immunohistochemistry Validation of IRE1p in Human Small Intestine Tissue. Immunohistochemical analysis of paraffin-embedded Human Small Intestine Tissue using anti-IRE1P antibody (orb1239523) at 2 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.

Immunofluorescence Validation of IRE1p in Rat Small Intestine Tissue. Immunofluorescent analysis of 4% paraformaldehyde-fixed Rat Small Intestine Tissue labeling IRE1p with orb1239523 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Immunohistochemistry Validation of IRE1p in Rat Small Intestine Tissue. Immunohistochemical analysis of paraffin-embedded Rat Small Intestine Tissue using anti-IRE1P antibody (orb1239523) at 2 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.

Induced Expression Validation of IRE1p in human umbilical vein endothelial cells (HUVECs) (Wang et al., 2019). IRE1p expression was examined by Western blot analysis with anti-IRE1p antibodies (orb1239523). IRE1p was increased in HUVEC cells treated with 10 µM 20 (S)‐PPD for 6 to 8 hours compared with control cells.