PA5-17160

antibody from Invitrogen Antibodies

Targeting: SERPINB5 maspin, PI5

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Antibody Data

Product number

PA5-17160

Provider

Invitrogen Antibodies

Product name

Maspin Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

It is not recommended to aliquot this antibody.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

125 µg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Maspin in extracts from HT-1080 and HT-29 cell lines, using Maspin polyclonal antibody (Product # PA5-17160).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Maspin Polyclonal Antibody(Product # PA5-17160) and a 42kDa band corresponding to Maspin was observed across cell lines and tissue extract tested except Caco-2, MDA-MB-231 and Mouse Adipose which is reported to be negative. Whole cell extracts (30 µg lysate) of MCF 10A (Lane 1), HaCaT (Lane 2), HeLa (Lane 3), Caco-2 (Lane 4), MDA-MB-231 (Lane 5) and tissue extracts of Mouse Adipose (Lane 6) and Mouse Mammary gland (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Maspin was performed using 70% confluent log phase MCF 10A 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 Maspin Polyclonal Antibody (Product # PA5-17160) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (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 cytoplasmic localization. Panel e represents negative cell line (MDA-MB-231) with no signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.