ProSci

IRE1p Antibody

Product Code:
 
PSI-3655
Product Group:
 
Primary Antibodies
Supplier:
 
ProSci
Host Type:
 
Rabbit
Antibody Isotype:
 
IgG
Antibody Clonality:
 
Polyclonal
Regulatory Status:
 
RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunofluorescence (IF)
  • Immunohistochemistry (IHC)
  • Western Blot (WB)
1 / 11
<strong>Figure 1 Western Blot Validation in Mouse A20 Cell Lysate</strong><br> Loading: 15 μg of lysates per lane. Antibodies: IRE1p 3655 (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.
2 / 11
<strong>Figure 2 KO Validation in HeLa Cells</strong><br> Loading: 10 μg of WT cell lysates (lane 1) or IRE1P KO cell lysates (lane 2). Antibodies: IRE1P 3655 (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.
3 / 11
<strong>Figure 3 Western Blot Validation in Human Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: IRE1p 3655 (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
4 / 11
<strong>Figure 4 Western Blot Validation in Rat Brain Tissue Lysate</strong><br> Loading: 15 μg of lysates per lane. Antibodies: IRE1p 3655 (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.
5 / 11
<strong>Figure 5 ImmunoFluor®scence Validation of IRE1p in Mouse A20 Cells</strong><br> ImmunoFluor®scent analysis of 4% paraformaldehyde-fixed A20 Cells labeling IRE1P with 3655 at 2 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
6 / 11
<strong>Figure 6 Immunocytochemistry Validation of IRE1p in Mouse A20 Cells</strong><br> Immunocytochemical analysis of A20 cells using anti-IRE1p antibody (3655) 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.
7 / 11
<strong>Figure 7 ImmunoFluor®scence Validation of IRE1p in Human Small Intestine Tissue</strong><br> ImmunoFluor®scent analysis of 4% paraformaldehyde-fixed Human Small Intestine Tissue labeling IRE1p with 3655 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
8 / 11
<strong>Figure 8 Immunohistochemistry Validation of IRE1p in Human Small Intestine Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded Human Small Intestine Tissue using anti-IRE1P antibody (3655) 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.
9 / 11
<strong>Figure 9 ImmunoFluor®scence Validation of IRE1p in Rat Small Intestine Tissue</strong><br> ImmunoFluor®scent analysis of 4% paraformaldehyde-fixed Rat Small Intestine Tissue labeling IRE1p with 3655 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
10 / 11
<strong>Figure 10 Immunohistochemistry Validation of IRE1p in Rat Small Intestine Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded Rat Small Intestine Tissue using anti-IRE1P antibody (3655) 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.
11 / 11
<strong>Figure 11 Induced Expression Validation of IRE1p in human umbilical vein endothelial cells (HUVECs) (Wang et al., 2019) </strong><br> IRE1p expression was examined by Western blot analysis with anti-IRE1p antibodies (3655). IRE1p was increased in HUVEC cells treated with 10 uM 20(S)?PPD for 6 to 8 hours compared with control cells.
❮❯

<strong>Figure 1 Western Blot Validation in Mouse A20 Cell Lysate</strong><br> Loading: 15 μg of lysates per lane. Antibodies: IRE1p 3655 (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.
<strong>Figure 2 KO Validation in HeLa Cells</strong><br> Loading: 10 μg of WT cell lysates (lane 1) or IRE1P KO cell lysates (lane 2). Antibodies: IRE1P 3655 (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.
<strong>Figure 3 Western Blot Validation in Human Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: IRE1p 3655 (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
<strong>Figure 4 Western Blot Validation in Rat Brain Tissue Lysate</strong><br> Loading: 15 μg of lysates per lane. Antibodies: IRE1p 3655 (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.
<strong>Figure 5 ImmunoFluor®scence Validation of IRE1p in Mouse A20 Cells</strong><br> ImmunoFluor®scent analysis of 4% paraformaldehyde-fixed A20 Cells labeling IRE1P with 3655 at 2 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
<strong>Figure 6 Immunocytochemistry Validation of IRE1p in Mouse A20 Cells</strong><br> Immunocytochemical analysis of A20 cells using anti-IRE1p antibody (3655) 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.
<strong>Figure 7 ImmunoFluor®scence Validation of IRE1p in Human Small Intestine Tissue</strong><br> ImmunoFluor®scent analysis of 4% paraformaldehyde-fixed Human Small Intestine Tissue labeling IRE1p with 3655 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
<strong>Figure 8 Immunohistochemistry Validation of IRE1p in Human Small Intestine Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded Human Small Intestine Tissue using anti-IRE1P antibody (3655) 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.
<strong>Figure 9 ImmunoFluor®scence Validation of IRE1p in Rat Small Intestine Tissue</strong><br> ImmunoFluor®scent analysis of 4% paraformaldehyde-fixed Rat Small Intestine Tissue labeling IRE1p with 3655 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
<strong>Figure 10 Immunohistochemistry Validation of IRE1p in Rat Small Intestine Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded Rat Small Intestine Tissue using anti-IRE1P antibody (3655) 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.
<strong>Figure 11 Induced Expression Validation of IRE1p in human umbilical vein endothelial cells (HUVECs) (Wang et al., 2019) </strong><br> IRE1p expression was examined by Western blot analysis with anti-IRE1p antibodies (3655). IRE1p was increased in HUVEC cells treated with 10 uM 20(S)?PPD for 6 to 8 hours compared with control cells.

No additional charges, what you see is what you pay! *

CodeSizePrice
PSI-3655-0.02mg0.02mg£150.00
Quantity:
PSI-3655-0.1mg0.1mg£449.00
Quantity:
Prices exclude any Taxes / VAT
How to order with us   Contact us
Stay in control of your spending. These prices have no additional charges, not even shipping!
* Rare exceptions are clearly labelled (only 0.14% of items!).
Multibuy discounts available! Contact us to find what you can save.
This product comes from: United States.
Typical lead time: 14-21 working days.
Contact us for more accurate information.
  • Further Information
  • Documents
  • References
  • Related Products
  • Show All

Further Information

Additional Names:
IRE1p Antibody: IRE1, IRE1P, IRE1a, hIRE1p, IRE1, Endoplasmic reticulum-to-nucleus signaling 1
Application Note:
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.
Background:
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.
Background References:
  • Little et al. Crit. Rev. Eukaryot. Gene Expr. 1994;4:1-18.
  • Lee. Methods 2005;35:373-81.
  • Bertolotti et al. Nat. Cell Biol. 2000;2:326-32.
  • Shen et al. Dev. Cell 2002;3:99-111.
Buffer:
IRE1p 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.
Immunogen:
Anti-IRE1p antibody (3655) 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.
ISOFORMS:
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.
NCBI Gene ID #:
2081
NCBI Official Name:
endoplasmic reticulum to nucleus signaling 1
NCBI Official Symbol:
ERN1
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 110kD

Observed: 110kD
Protein Accession #:
O75460
Protein GI Number:
193806335
Purification:
IRE1p Antibody is affinity chromatography purified via peptide column.
Research Area:
Signal Transduction
Swissprot #:
O75460
User NOte:
Optimal dilutions for each application to be determined by the researcher.
VALIDATION:

KO validation (Figure 2): Anti-IRE1p antibody (3655) specificity was further verified by IRE1P specific knockout. IRE1p signal was disrupted in IRE1p knockout HeLa cells in comparison with that in control HeLa cells.

Induced Expression Validation (Figure 11): IRE1p expression detected by anit-IRE1p antibodies was up-regulated by 20(S)?PPD treatment.

References

  1. Wang et al. 20(S)-protopanaxadiol induces apoptosis in human umbilical vein endothelial cells by activating the PERK-eIF2alpha-ATF4 signaling pathway. J Cell Biochem. 2019;120(4):5085-5096.PMID: 30259568
  2. 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: