PA5-101662

antibody from Invitrogen Antibodies

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

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Antibody Data

Product number

PA5-101662

Provider

Invitrogen Antibodies

Product name

MBTPS1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Antibody detects endogenous levels of total MBTPS1.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

1 mg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of MBTPS1 in HeLa cell lysate (Lane 1: treated with the antigen-specific peptide). Samples were incubated with MBTPS1 polyclonal antibody (Product # PA5-101662).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of MBTPS1 in Jurkat/COLO205 cells. Samples were incubated with MBTPS1 polyclonal antibody (Product # PA5-101662).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-MBTPS1 Polyclonal Antibody (Product # PA5-101662) 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) 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). Increased expression was observed after PTI treatment. Two non specific bands around 40 and 60 kDa were observed.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of MBTPS1 in HeLa cells. Samples were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100, blocked with 10% serum (45 min at 25°C) incubated with MBTPS1 polyclonal antibody (Product # PA5-101662) using a dilution of 1:200 (1 hr, 37°C), and followed by goat anti-rabbit IgG Alexa Fluor 594 at a dilution of 1:600.

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 # PA5-101662) at 1:200 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.