Anti-ATF6 antibody

• WB

Supplier Data

Western blot - Anti-ATF6 antibody (AB37149)

1 h incubation at RT in 5% NFDM/TBST.

All lanes:

Western blot - Anti-ATF6 antibody (ab37149) at 0.5 µg/mL

Lane 1:

Wild type HeLa cells at 15 µg

Lane 2:

ATF6 knockout HeLa cells at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG HRP conjugate at 1/10000 dilution

Predicted band size: 74 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF6 antibody (AB37149)

Immunohistochemical analysis of paraffin-embedded Human testis tissue using ab37149 at 5µg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1h at RT. Antigen retrieval was by heat mediation with a citrate buffer (pH 6.0). Samples were incubated with primary antibody overnight at 4°C. Goat anti-rabbit IgG H&L (HRP) at 1/250 dilution was used as a secondary antibody. Counter stained with Hematoxylin.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF6 antibody (AB37149)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue using ab37149 at 5µg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1h at RT. Antigen retrieval was by heat mediation with a citrate buffer (pH 6.0). Samples were incubated with primary antibody overnight at 4°C. Goat anti-rabbit IgG H&L (HRP) at 1/250 dilution was used as a secondary antibody. Counter stained with Hematoxylin.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF6 antibody (AB37149)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue using ab37149 at 5µg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1h at RT. Antigen retrieval was by heat mediation with a citrate buffer (pH 6.0). Samples were incubated with primary antibody overnight at 4°C. Goat anti-rabbit IgG H&L (HRP) at 1/250 dilution was used as a secondary antibody. Counter stained with Hematoxylin.

• WB

Supplier Data

Western blot - Anti-ATF6 antibody (AB37149)

1 h incubation at RT in 5% NFDM/TBST.

All lanes:

Western blot - Anti-ATF6 antibody (ab37149) at 1 µg/mL

Lane 1:

HeLa cell lysates at 15 µg

Lane 2:

HepG2 cell lysates at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG HRP conjugate at 1/10000 dilution

Predicted band size: 74 kDa

false

• WB

Supplier Data

Western blot - Anti-ATF6 antibody (AB37149)

Blocking buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ATF6 antibody (ab37149) at 1 µg/mL

Lane 1:

Mouse Heart Lysate at 15 µg

Lane 2:

Mouse Skeletal Muscle Lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG HRP Conjugate at 1/10000 dilution

Predicted band size: 74 kDa

false

• WB

Supplier Data

Western blot - Anti-ATF6 antibody (AB37149)

Blocking buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ATF6 antibody (ab37149) at 1 µg/mL

All lanes:

Rat Kidney Lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG HRP Conjugate at 1/10000 dilution

Predicted band size: 74 kDa

false

• WB

CiteAb

Western blot - Anti-ATF6 antibody (AB37149)

Western Blotting using Anti-ATF6 antibody, ab37149. Publication image from Yang, Q. et al., 2017, Nat Commun, 29176575. Legend direct from paper.

PERK and MKK4 are required to activate the p38 MAPK–CMA pathway under ER stress. a siRNAs effectively knockdown PERK, IRE1, and ATF6. SN4741 cells were transfected with siRNAs for 72 h. b, c Knockdown of PERK abolishes the activation of CMA by ER stress. SN4741 cells were transfected with siRNAs for 60 h, then treated with Tg (0.3 µM) or Tu (3 µg/ml) for 12 h, and assessed for endogenous CMA substrate MEF2D level b and RNase A uptake c. d Knockdown of PERK reduces p-LAMP2A. SN4741 cells were transfected siRNAs for 69 h, and then treated with Tg for 2 h. e MKK3 and 6 are not involved in ER stress-induced CMA. SN4741 cells were transfected with si-Control, si-MKK3, si-MKK6, and si-MKK3 as well as 6 (MKK3/6) for 72 h. Total proteins were blotted with MKK3 and MKK6 antibodies (left). SN4741 cells were transfected for 70 h and exposed to Tg or Tu for 2 h for p-p38 MAPK (top right) or transfected for 60 h and exposed to Tg or Tu for 12 h for MEF2D (bottom right). f, g Knockdown of MKK4 attenuates activation of p38 MAPK and phosphorylation of LAMP2A induced by ER stress. SN4741 cells were transfected with si-Control and si-MKK4 for 70 h, and then treated with Tg or Tu for 2 h (f) or transfected for 69 h and stressed for 3 h (g). h, i Knockdown of MKK4 reverses the reduction of MEF2D induced by ER stress. SN4741 cells were transfected with si-Control and si-MKK4 for 60 h, treated with Tg or Tu for 12 h, and assessed for MEF2D level (h) and RNase A uptake (i). j Knockdown of PERK reverses the activation of MKK4 and p38 MAPK by ER stress. SN4741 cells were transfected with si-Control and si-PERK for 70 h, and then treated with Tg or Tu for 2 h. Quantifications are shown for b, e, and h (n = 3. All values are s.d. (one-way ANOVA with Turkey test). **p < 0.005 vs. control and ##p < 0.005 vs. corresponding challenge alone)

false