PA5-27251

antibody from Invitrogen Antibodies

Targeting: S100A10 42C, ANX2LG, CAL1L, CLP11, P11

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-27251

Provider

Invitrogen Antibodies

Product name

S100A10 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Recommended positive controls: 293T, A431, H1299, HeLa.

Concentration

1 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using S100A10 Polyclonal Antibody (Product # PA5-27251). Sample (30 µg of whole cell lysate). Lane A: A431 .15% SDS PAGE. S100A10 Polyclonal Antibody (Product # PA5-27251) diluted at 1:1,000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-S100A10 Polyclonal Antibody (Product # PA5-27251) and a 11kDa band corresponding to Protein S100-A10 was observed across the panel tested except SW480 which is reported to be negative. Whole cell extracts (30 µg lysate) of HaCaT (Lane 1), HeLa (Lane 2), PANC-1 (Lane 3), HT-29 (Lane 4), SW480 (Lane 5), NIH:OVCAR-3 (Lane 6), NIH:OVCAR-3 (treated with retinoic acid, 5uM for 6 days) (Lane 7) 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 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 Protein S100-A10 was performed using 70% confluent log phase HT-29 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 15 minutes at room temperature. The cells were labeled with S100A10 Polyclonal Antibody (Product # PA5-27251) at 1:200 dilution 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), (1:3000 dilution), 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 merged image for SW480 cells showing no staining for S100A10. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded human gastric cancer, using S100A10 (Product # PA5-27251) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.