Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Immunohistochemistry [2]
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Validation data
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- Product number
- MA5-26658 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- S100A6 Monoclonal Antibody (OTI5A10)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- OTI5A10
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of S100A6 in HEK293T cells in untransfected (Left lane) and transfected (Right lane) samples using 5 µg per lane. The samples were separated by SDS-PAGE and probed with S100A6 (Product # MA5-26658) monoclonal antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-S100A6 Monoclonal Antibody (OTI5A10) (Product # MA5-26658) and a 10kDa band corresponding to Protein S100-A6 was observed across. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), PC-3 (Lane 3), U-87 MG (Lane 4), HEK-293 (Lane 5), K-562 (Lane 6) and LNCaP (Lane 7) were electrophoresed using Novex™ 16% Tricine Protein Gel (Product # EC6695BOX). 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 (1:2000) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Band of interest was not observed in HEK-293, K-562 and LNCaP which are reported to be either negative or express S100A6 at very low levels.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of S100A6 was performed using 70% confluent log phase U-87 MG 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 S100A6 Monoclonal Antibody (OTI5A10) (Product # MA5-26658) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominant cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7C1115LZR).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded carcinoma of human lung tissue. To expose target proteins, 1 mM EDTA in 10mM Tris, pH8.5, 120°C for 3min was used. Following antigen retrieval, tissues were probed with a S100A6 monoclonal antibody (Product # MA5-26658).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded carcinoma of human pancreas tissue. To expose target proteins, 1 mM EDTA in 10mM Tris, pH8.5, 120°C for 3min was used. Following antigen retrieval, tissues were probed with a S100A6 monoclonal antibody (Product # MA5-26658).