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Active Activating Transcription Factor 6 (ATF6)

Processed cyclic AMP-dependent transcription factor ATF-6 alpha

ACTIVITY TEST

Activating Transcription Factor 6 (ATF6) is a key protein in the endoplasmic reticulum (ER) stress response. It is a transmembrane protein located in the ER membrane. Under normal conditions, ATF6 is bound to Heat Shock 70kDa Protein 5 (HSPA5), also known as GRP78, and remains inactive.When ER stress occurs due to the accumulation of unfolded or misfolded proteins in the ER lumen, HSPA5 preferentially binds to these abnormal proteins, releasing ATF6. The released ATF6 then translocates to the Golgi apparatus, where it is cleaved by proteases. The cleaved ATF6 fragment enters the nucleus and activates the transcription of genes involved in the unfolded protein response, helping the cell to restore ER homeostasis.Thus a functional ELISA assay was conducted to detect the interaction of recombinant mouse ATF6 and recombinant mouse HSPA5. Briefly, ATF6 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to HSPA5-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-ATF6 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630nm immediately. The binding activity of recombinant mouse ATF6 and recombinant mouse HSPA5 was shown in Figure 1, the EC50 for this effect is 0.031ug/mL.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Catalog No. Related products for research use of Mus musculus (Mouse) Organism species Applications (RESEARCH USE ONLY!)
APB393Mu01 Active Activating Transcription Factor 6 (ATF6) Cell culture; Activity Assays.
RPB393Mu01 Recombinant Activating Transcription Factor 6 (ATF6) Positive Control; Immunogen; SDS-PAGE; WB.
RPB393Mu02 Recombinant Activating Transcription Factor 6 (ATF6) Positive Control; Immunogen; SDS-PAGE; WB.
PAB393Mu01 Polyclonal Antibody to Activating Transcription Factor 6 (ATF6) WB; IHC
PAB393Mu02 Polyclonal Antibody to Activating Transcription Factor 6 (ATF6) WB; IHC; ICC; IP.
SEB393Mu ELISA Kit for Activating Transcription Factor 6 (ATF6) Enzyme-linked immunosorbent assay for Antigen Detection.
LMB393Mu Multiplex Assay Kit for Activating Transcription Factor 6 (ATF6) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.