PA5-13921

antibody from Invitrogen Antibodies

Targeting: ERN2 IRE1b

Western blot (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-13921

Provider

Invitrogen Antibodies

Product name

ERN2 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

400 µL

Concentration

2 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

IRE1β negatively regulates IRE1α signaling in response to endoplasmic reticulum stress.

Grey MJ, Cloots E, Simpson MS, LeDuc N, Serebrenik YV, De Luca H, De Sutter D, Luong P, Thiagarajah JR, Paton AW, Paton JC, Seeliger MA, Eyckerman S, Janssens S, Lencer WI
The Journal of cell biology 2020 Feb 3;219(2)

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 6. IRE1beta inhibits IRE1alpha endonuclease activity in vitro. (A) Samples of affinity-purified full-length IRE1alpha and IRE1beta were separated on SDS-PAGE and stained with Coomassie blue or assayed by immunoblot with anti-IRE1alpha, anti-IRE1beta, or anti-Myc antibodies. (B) In vitro endonuclease activity was assayed by monitoring cleavage of fluorescent reporter substrate (10 nM) over time for indicated concentrations of (top, purple) IRE1alpha or (bottom) IRE1beta. Bottom, inset: Endonuclease activity monitored over 24 h for 250 nM IRE1beta (light green) or buffer control (black circles). (C) Steady-state kinetics were assayed by measuring progress curves for 10 nM IRE1alpha or 10 nM IRE1alpha + 50 nM IRE1beta as a function of reporter substrate concentration. Kinetic data are plotted as initial reaction velocity versus substrate concentration. Data were measured for two independent preparations of purified protein, with symbols representing mean +- SEM for three independent measures for preparation 1 (Prep#1) or values from a single measurement for preparation 2 (Prep#2). Solid lines represent best fit of a noncompetitive inhibitor model to the kinetic data. (D) Top: Gel filtration chromatogram for lysate of HEK293 cells expressing IRE1alpha-mCherry. A high-molecular-weight (HMW) fraction (indicated by filled symbol in top panel) was incubated with either buffer (purple traces) or purified IRE1beta (100 nM, light blue traces), reinjected on gel filtration, and as