Anti-IRE1 (phospho S724) antibody (ab48187)
Key features and details
- Rabbit polyclonal to IRE1 (phospho S724)
- Suitable for: WB, ELISA, IHC-P
- Reacts with: Mouse, Human, African green monkey
- Isotype: IgG
Overview
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Product name
Anti-IRE1 (phospho S724) antibody
See all IRE1 primary antibodies -
Description
Rabbit polyclonal to IRE1 (phospho S724) -
Host species
Rabbit -
Specificity
ab48187 detects IRE1 alpha, phosphorylated (serine 724) protein. -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P HumanWB MouseHumanAfrican green monkey
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Immunogen
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Positive control
- WB: HeLa, Min6, COS-7 cells. IHC-P: Human spleen tissue.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.40
Preservative: 0.025% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-IRE1 (phospho S724) antibody (ab48187)All lanes : Anti-IRE1 (phospho S724) antibody (ab48187) at 1/2000 dilution
Lane 1 : HeLa cells 30 nM Calyculin A: AP-buffer
Lane 2 : HeLa cells 30 nM Calyculin A: Alkaline Phosphatase
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : infrared (IR)-labelled goat anti-rabbit (green) antibody and IR-labelled goat anti-mouse (red) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 110 kDaThe blots were produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membranes were blocked for an hour. Membrane 2 was incubated with alkaline phosphatase (AP; 100 U per mL) for one hour, whilst membrane 1 was treated with AP-buffer only, before being incubated with ab48187 (rabbit anti-IRE1 antibody diluted 1:2000) and loading control ab125247 (mouse anti-GAPDH antibody; diluted 1:10,000) for 24 hours at 4°C. Antibody binding was detected using infrared (IR)-labelled goat anti-rabbit (green) antibody and IR-labelled goat anti-mouse (red) at 1:10,000 dilutions for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRE1 (phospho S724) antibody (ab48187)Paraffin-embedded human spleen tissue stained for IRE1 using ab48187 at 1/300 dilution for 1 hour at room temperature in immunohistochemical analysis. DAB staining. Counter stained with hematoxylin.
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ELISA - Anti-IRE1 (phospho S724) antibody (ab48187)Serially diluted ab48187 was bound to immobilised Phospho peptide (133861) - or Control peptide (1 microgram x mL-1). The antibody was detected by HRP-labelled goat anti-rabbit IgG (ab97080; diluted 50000 times) and signal was developed with TMB substrate.
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Western blot - Anti-IRE1 (phospho S724) antibody (ab48187)All lanes : Anti-IRE1 (phospho S724) antibody (ab48187) at 1/1000 dilution
Lane 1 : Min6 cells untreated
Lane 2 : Min6 cells treated with glucose for 3 hours at 5 mM
Lane 3 : Min6 cells treated with glucose for 3 hours at 20 mM
Developed using the ECL technique.
Predicted band size: 110 kDa
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Western blot - Anti-IRE1 (phospho S724) antibody (ab48187)All lanes : Anti-IRE1 (phospho S724) antibody (ab48187) at 1/2000 dilution
Lane 1 : Cell lysate prepared from COS-7 Untransfected cells
Lane 2 : Cell lysate prepared from COS-7 cells expressing wild type IRE1 alpha
Lane 3 : Cell lysate prepared from COS-7 cells expressing kinase-dead IRE1 alpha
Predicted band size: 110 kDa
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Western blot - Anti-IRE1 (phospho S724) antibody (ab48187)All lanes : Anti-IRE1 (phospho S724) antibody (ab48187) at 2 µg/ml
Lane 1 : Untreated HeLa cells (Batch 1 ab48187)
Lane 2 : DTT-treated HeLa cells (Batch 1 ab48187)
Lane 3 : Untreated HeLa cells (Batch 2 ab48187)
Lane 4 : DTT-treated HeLa cells (Batch 2 ab48187)
Secondary
All lanes : Anti-Rabbit IgG HRP
Predicted band size: 110 kDaHeLa cells were treated (+) or untreated (-) with 10 mM DTT for 60 min to activate the UPR. Total protein was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% BSA in TBST.
