Anti-IRE1p ERN1 Antibody

Product Name

Anti-IRE1p ERN1 Antibody

View all IRE1 alpha Antibodies

SKU/Catalog Number

A00683

Size

0.1 mg

Form

Liquid

Description

Boster Bio Anti-IRE1p ERN1 Antibody (Catalog # A00683). Tested in ELISA, WB, ICC, IF, IHC-P applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

IRE1p antibody can be stored at 4°C for three months and -20°C, stable for up to one year. Avoid repeated freeze-thaw cycles. Antibodies should not be exposed to prolonged high temperatures.

Cite This Product

Anti-IRE1p ERN1 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00683)

Host

Rabbit

Contents

IRE1p Antibody is supplied in PBS containing 0.02% sodium azide.

Clonality

Polyclonal

Isotype

IgG

Immunogen

Anti-IRE1p antibody 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.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

SHBG antibody is human, mouse, and rat reactive. At least three isoforms of SHBG are known to exist; this antibody will detect all three isoforms.

Reactive Species

A00683 is reactive to ERN1 in Human, Mouse, Rat

Applications

A00683 is guaranteed for ELISA, IF, IHC-P, ICC, WB Boster Guarantee

Observed Molecular Weight

68 kDa

Calculated molecular weight

109.735kDa

Background of IRE1 alpha

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.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

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. Optimal dilutions for each application should be determined by the researcher.

Validation Images & Assay Conditions

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Western Blot Validation in Mouse A20 Cell Lysate
Loading: 15 μg of lysates per lane. Antibodies: IRE1p A00683 (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.

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KO Validation in HeLa Cells
Loading: 10 μg of WT cell lysates (lane 1) or IRE1P KO cell lysates (lane 2).Antibodies: IRE1P A00683 (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.

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Western Blot Validation in Human Cell Lines
Loading: 15 μg of lysates per lane. Antibodies: IRE1p A00683 (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

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Western Blot Validation in Rat Brain Tissue Lysate
Loading: 15 μg of lysates per lane. Antibodies: IRE1p A00683 (A: 0.5 μg/mL

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Immunofluorescence Validation of IRE1p in Mouse A20 Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed A20 Cells labeling IRE1P with A00683 at 2 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).

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Immunocytochemistry Validation of IRE1p in Mouse A20 Cells
Immunocytochemical analysis of A20 cells using anti-IRE1p antibody (A00683) 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.

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Immunofluorescence Validation of IRE1p in Human Small Intestine Tissue
Immunofluorescent analysis of 4% paraformaldehyde-fixed Human Small Intestine Tissue labeling IRE1p with A00683 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

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Immunofluorescence Validation of IRE1p in Rat Small Intestine Tissue
Immunofluorescent analysis of 4% paraformaldehyde-fixed Rat Small Intestine Tissue labeling IRE1p with A00683 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

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Induced Expression Validation of IRE1p in human umbilical vein endothelial cells (HUVECs) (Wang et al.

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Immunohistochemistry Validation of IRE1p in Human Small Intestine Tissue
Immunohistochemical analysis of paraffin-embedded Human Small Intestine Tissue using anti-IRE1P antibody (A00683) 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.

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Immunohistochemistry Validation of IRE1p in Rat Small Intestine Tissue
Immunohistochemical analysis of paraffin-embedded Rat Small Intestine Tissue using anti-IRE1P antibody (A00683) 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.