IRE1p Antibody

Signal Transduction

WB: 0.5-2 μ g/mL; ICC: 1 μ g/mL; IF: 2-20 μ g/mL; IHC: 2 μ g/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.

Cat. No. 17-208 - A-20 Cell Slide

Predicted: 110kD

Observed: 110kD

Human IRE1p has 2 isoforms, including isoform 1 (977aa, 110kD) and isoform 2 (70aa, 6.6kD). Mouse IRE1p also has two isoforms, including isoform 1 (977aa, 110kD) and isoform 2 (408aa, 45kD). Rat IRE1p has one isoform (965aa, 109kD). 3655 can detect human, mouse and rat.

Polyclonal

Unconjugated

1 mg/mL

Cat. No. 3655P – IRE1p Peptide

None

O75460

2081

ERN1

Homo sapiens

IRE1p Antibody: Accumulation of malfolded proteins in the endoplasmic reticulum (ER) activates the unfolded protein response (UPR) and the upregulation of the ER molecular chaperones GRP78 and GRP 94. These proteins are normally bound to ER transmembrane proteins such as IRE1p and ATF6 but ER stress causes their dissociation. This allows IRE1p, a serine-threonine protein kinase to transduce the unfolded protein signal from the ER to the nucleus. IRE1p also has an endoribonuclease activity that is required to splice X-box binding protein (XBP1) mRNA converting it to a potent UPR transcriptional activation. Depletion of IRE1p through the expression of a dominant negative form of IRE1p has no effect on transfected cells, but cell death via apoptosis occurs under stress conditions that cause unfolded proteins to accumulate in the ER. Two alternatively spliced transcript variants encoding different isoforms have been found for this gene.

Lee. Methods 2005; 35:373-81.

Shen et al. Dev. Cell 2002; 3:99-111.

Zhang et al. 20(S)-protopanaxadiol activates ER stress to induce cell apoptosis in human umbilical vein endothelial cells. Journal of Shanghai University of Traditional Chinese Medicine 2015; 29(3):70-75.PMID:

Figure 2 KO Validation in HeLa Cells
Loading: 10 μ g of WT cell lysates (lane 1) or IRE1P KO cell lysates (lane 2).Antibodies: IRE1P 3655 (0.5 μ g/mL) and beta-actin (1 μ g/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Figure 4 Western Blot Validation in Rat Brain Tissue Lysate
Loading: 15 μ g of lysates per lane.Antibodies: IRE1p 3655 (A: 0.5 μ g/mL, B: 1 μ g/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Figure 6 Immunocytochemistry Validation of IRE1p in Mouse A20 Cells
Immunocytochemical analysis of A20 cells using anti-IRE1p antibody (3655) at 1 μ g/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 8 Immunohistochemistry Validation of IRE1p in Human Small Intestine Tissue
Immunohistochemical analysis of paraffin-embedded Human Small Intestine Tissue using anti-IRE1P antibody (3655) at 2 μ g/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.

Figure 10 Immunohistochemistry Validation of IRE1p in Rat Small Intestine Tissue
Immunohistochemical analysis of paraffin-embedded Rat Small Intestine Tissue using anti-IRE1P antibody (3655) at 2 μ g/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.