Phospho-IRE1 (S724) Recombinant Rabbit Monoclonal Antibody [PSH03-35]
cat.: HA721980
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse |
| Applications: | WB, IF-Cell, IHC-P, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH03-35 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 110 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser724 of human IRE1. |
| Positive control: | HeLa starved for 3 hours then treated with 100nM Calyculin A for 30 minutes cell lysate, Jurkat treated with 100nM Calyculin A for 30 minutes cell lysate, Jurkat cells treated with 100nM Calyculin A for 30 minutes, human pancreas tissue, HeLa cells treated with 100nM Calyculin A for 30 minutes. |
| Subcellular location: | Endoplasmic reticulum membrane; nucleus. |
| Recommended Dilutions:
WB IF-Cell IHC-P FC |
1:1,000 1:500 1:500 1:1,000 |
| Uniprot #: | SwissProt: O75460 Human | Q9EQY0 Mouse |
| Alternative names: | Endoplasmic reticulum (ER) to nucleus signalling 1 Endoplasmic reticulum to nucleus signaling 1 Endoplasmic reticulum-to-nucleus signaling 1 Endoribonuclease ER to nucleus signaling 1 ERN 1 Ern1 ERN1_HUMAN hIRE 1p hIRE1p Inositol requiring 1 Inositol requiring 1, S. cerevisiae, homolog of Inositol requiring enzyme 1, S. cerevisiae, homolog of Inositol requiring protein 1 inositol-requiring enzyme 1 Inositol-requiring protein 1 IRE 1 IRE 1a IRE 1P Ire1 alpha Ire1-alpha IRE1a Ire1alpha IRE1P MGC163277 MGC163279 Protein kinase/endoribonuclease RGD1559716 Serine/threonine protein kinase/endoribonuclease IRE1 |
Images
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Fig1:
Western blot analysis of Phospho-IRE1 (S724) on different lysates with Rabbit anti-Phospho-IRE1 (S724) antibody (HA721980) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa starved for 3 hours then treated with 100nM Calyculin A for 30 minutes cell lysate Lane 3: Jurkat cell lysate Lane 4: Jurkat treated with 100nM Calyculin A for 30 minutes cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 110 kDa Observed band size: 110 kDa Exposure time: 5 minutes 10 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721980) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of Jurkat cells treated with 100nM Calyculin A for 30 minutes labeling Phospho-IRE1 (S724) with Rabbit anti-Phospho-IRE1 (S724) antibody (HA721980) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-IRE1 (S724) antibody (HA721980) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig3:
Immunohistochemical analysis of paraffin-embedded human pancreas tissue with Rabbit anti-Phospho-IRE1 (S724) antibody (HA721980) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721980) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig4:
Flow cytometric analysis of HeLa cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-IRE1 (S724). Cells were fixed and permeabilized. Then stained with the primary antibody (HA721980, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.