Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunohistochemistry [1]
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- Product number
- MA5-16380 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- MSH6 Monoclonal Antibody (SP93)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- Heat-mediated antigen retrieval is recommended prior to staining, using a 1mM EDTA buffer, pH 8.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 30 min at room temperature. A suggested positive control is rectal carcinoma.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- SP93
- Vial size
- 500 µL
- Storage
- 4° C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- CRISPR-Cas9 mediated genome editing ofMSH6 (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR1065222_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of MSH6 was performed by loading 30 µg of HeLa wild type (Lane 1), HeLa Cas9 (Lane 2) and HeLa Cas9 cells transduced with MSH6 Lentiviral sgRNA (Lane 3) modified whole cell extracts. The samples were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-MSH6 Monoclonal Antibody (SP93) (Product # MA5-16380) using 1:1000 dilution and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036 1:5000 dilution).Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). A reduced signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific toMSH6 (Fig (b)). Uncharacterized band was observed in all the samples at ~40 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-MSH6 Monoclonal Antibody (SP93) (Product # MA5-16380) and a 160kDa band corresponding to MSH6 was observed in all the tested cell models, except Jurkat and LNCaP which are reported to be negative. This target undergoes heavy phosphorylation and multiple bands between 120-160kDa can be expected. Modified whole cell lysate (1% SDS) (30ug lysate) of SH-SY5Y (Lane 1), K-562 (Lane 2), Jurkat (Lane 3), HeLa (Lane 4), LNCaP (Lane 5), PC-3 (Lane 6), HCT 116 (Lane 7) and A549 (Lane 8) were electrophoresed using NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX). 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).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of A431 Cells using anti-MSH6 Monoclonal Antibody (Product # MA5-16380). The recommened dilution for this antibody in western blot applications is 1:100.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of MSH6 using anti-MSH6 Monoclonal Antibody (Product # MA5-16380) in Rectal Cancer Tissue. The recommened dilution for this antibody in immunohistochemistry applications is 1:100.