PA5-109948

antibody from Invitrogen Antibodies

Targeting: MBTPS1 KIAA0091, PCSK8, S1P, SKI-1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-109948

Provider

Invitrogen Antibodies

Product name

MBTPS1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

3.46 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of MBTPS1 in K-562, mouse pancreas. Samples were incubated in polyclonal MBTPS1 antibody (Product # PA5-109948) using a dilution of 1:1000, followed by HRP Goat Anti-Rabbit IgG (H+L) at a dilution of 1:10000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-MBTPS1 Polyclonal Antibody (Product # PA5-109948) and a ~130 kDa band corresponding to Membrane-bound transcription factor site-1 protease was observed across cell lines tested . Membrane enriched extracts (50 µg lysate) of SK-O-V3 (Lane 1), SK-O-V3 treated with PTI (Lane 2), A549 (Lane 3), A549 treated with PTI (Lane 4), MCF7 (Lane 5), MCF7 treated with PTI (Lane 6), HeLa (Lane 7), K-562 (Lane 8), u-87MG (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580). A non specific band was observed around 35 kDa. Increased expression was observed in cell lines post PTI treatment.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Membrane-bound transcription factor site-1 protease was performed using 70% confluent log phase MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with MBTPS1 Polyclonal Antibody (Product # PA5-109948) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing endoplasmic reticulum and golgi apparatus localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.