Anti-SEC61A antibody [EPR14379]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SEC61A antibody [EPR14379] (AB183046)

Immunohistochemical analysis of paraffin-embedded Human infiltrating duct carcinoma of breast tissue labeling SEC61A with ab183046 at 1/100 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-SEC61A antibody [EPR14379] (AB183046)

Western blot analysis of SEC61A in Daudi cell lysate immunoprecipitated using ab183046 at 1/50 dilution (Lane 1). Lane 2 : Negative control.

Secondary antibody : Anti-Rabbit IgG (HRP) specific to the non-reduced form of IgG at 1/1500 dilution.

All lanes:

Immunoprecipitation - Anti-SEC61A antibody [EPR14379] (ab183046)

Predicted band size: 52 kDa

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• WB

Supplier Data

Western blot - Anti-SEC61A antibody [EPR14379] (AB183046)

All lanes:

Western blot - Anti-SEC61A antibody [EPR14379] (ab183046) at 1/5000 dilution

Lane 1:

Human fetal brain lysate at 20 µg

Lane 2:

Daudi cell lysate at 20 µg

Lane 3:

A431 cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 52 kDa

Observed band size: 49 kDa

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• WB

CiteAb

Western blot - Anti-SEC61A antibody [EPR14379] (AB183046)

SEC61A western blot using anti-SEC61A antibody [EPR14379] ab183046. Publication image and figure legend from Puri, C., Vicinanza, M., et al., 2018, Dev Cell, PubMed 29634932.

ab183046 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab183046 please see the product overview.

TfR Is Recruited to LC3 Vesicles in Starvation Conditions(A) HeLa cells transfected with GFP-LC3, were loaded with transferrin Alexa 555 for 1 hr in Hank's balanced salt solution or full medium and chased for 15 min. The Tf/LC3 and LC3/Tf signal overlaps are shown. Data are means ± SEM, n = 3; two-tailed paired t test, ***p < 0.001.(B) HeLa cells treated as in (A) were processed for immunogold labeling on cryosections. Arrows specify autophagosome double membranes, black outer, white inner.(C) HeLa cells starved for 1 hr and loaded with anti-TfR antibody were processed for pre-embedding electron microscopy.(D) HeLa cells treated with ATG2A/B siRNA (left) were processed as in (C). HeLa cells transfected for Flag-ATG4BC74A were loaded with Tf and processed as in (B). LD, lipid droplet. Arrows specify autophagosome double membranes as in (B).(E) HeLa cells treated with control or ATG7 siRNA and/or transfected with ATG7 were processed for immunoblot and the TfR levels measured. Data are means ± SD, n = 3; two-tailed paired t test, ***p < 0.001.(F) HeLa cells treated with control or RAB11A or WIPI2 siRNAs were processed as in (E). Data are means ± SD, n = 3; two-tailed paired t test, **p < 0.01, ***p < 0.001.(G) HeLa cells starved for 1 hr, loaded with Ferrofluid-Tf Alexa 488 for 1 hr and chased for 15 min in starvation medium. The cells were then fragmented and the membranes containing Ferrofluid-Tf488 (bound) or not containing Ferrofluid-Tf488 (unbound) were separated and processed for immunoblot (see Figure S5G).(H) HeLa cells transfected with control or ATG2A/B siRNA were processed as in (G). The amount of LC3II in bound fraction in ATG2 knockdown condition is expressed as percentage of control. Data are means ± SEM, n = 3.(I) HeLa cells treated with control or ULK1 siRNA and processed as in (G). The LDH activity was measured in the bound fraction. Data are means ± SEM, n = 3; two-tailed paired t test, ***p < 0.001.

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