Immunofluorescence analysis of SEC61B was performed using 70% confluent log phase MDCK cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with SEC61B Rabbit Polyclonal Antibody (PA3015) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Dog, Human, Mouse, Pig, Rat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues M(1) P G P T P S G T N V G S S G R S P S K(20) of canine sec61 beta.|
|Storage buffer||whole serum|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:20,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunoprecipitation (IP)||See 1 publications below|
PA3-015 detects human, canine and porcine, and rat sec61 beta from microsomes and shows no cross-reactivity with other sec61 isoforms.
PA3-015 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~14 kDa protein representing sec61 beta from pig microsomes.
The PA3-015 immunogen is a synthetic peptide corresponding to residues M(1) P G P T P S G T N V G S S G R S P S K(20) of canine sec61 beta. This sequence is completely conserved between human, mouse, and canine.
The Sec61 complex is responsible for translocation of proteins across the rough endoplasmic reticulum membrane. Sec61 is composed of three subunits: alpha, beta, and gamma; that associate to form a heterotrimer. Multiple heterotrimers associate forming the ER translocon. Sec61 alpha is a polytopic protein that has ten membrane-spanning loops.
Sec61 has been found to be tightly associated with membrane bound ribosomes, either directly or through adaptor proteins, and is required for assembly of membrane and secretory proteins.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Characterization of binding of LARP6 to the 5' stem-loop of collagen mRNAs: implications for synthesis of type I collagen.
PA3-015 was used in immunoprecipitation and western blot to investigate the contribution of the unique sequence of LARP6 to type I collagen biosynthesis
|Stefanovic L,Longo L,Zhang Y,Stefanovic B||RNA biology (11:1386)||2015|