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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [4]
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- Product number
- MA1-41246 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- MLH1 Monoclonal Antibody (164C819)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- This peptide sequence is 93% homologous to mouse and dog MLH1. Suggested positive control: Daudi cell lysate.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 164C819
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of MLH1 inDaudicell lysate using a MutL protein homolog 1 monoclonal antibody (Product # MA1-41246) at 2 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of MLH1 in Daudi cell lysate. Samples were incubated in MLH1 monoclonal antibody (Product # MA1-41246) using a dilution of 2 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockout of MLH1 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR1027533_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of MLH1 was performed by loading 30 µg of HeLa Wild Type (Lane 1), HeLa Cas9 (Lane 2) and Hela MLH1 KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 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 Anti-MLH1 Monoclonal Antibody (164C819) (Product # MA1-41246, 2 µg/mL dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to MLH1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-MLH-1 Mouse Monoclonal Antibody (Product # MA1-41246) and a 84 kDa band corresponding to MLH-1 was observed in cell lines tested except HCT 116 and DU 145. Modified whole cell extracts (30 µg lysate) of HeLa (Lane 1), A431 (Lane 2), SW480 (Lane 3), HCT 116 (Lane 4), PC3 (Lane 5), DU 145 (Lane 6) and Raji (Lane 7) were electrophoresed using Novex® 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 Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
