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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Flow cytometry [1]
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Validation data
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- Product number
- MA5-28133 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Talin Monoclonal Antibody (TA205)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Mouse anti Human Talin antibody, clone TA205 recognizes human talin, a approximately 225 kDa cytoskeletal protein.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- TA205
- Vial size
- 200 µg
- Concentration
- 1 mg/mL
- Storage
- 4°C or -20°C if preferred
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Talin 1 Polyclonal Antibody (Product # MA5-28133) and a 269 kDa band corresponding to Talin-1 was observed across the cell lines tested. Whole cell extracts of A549 (Lane 1), K-562 (Lane 2), MDA-MB-231 (Lane 3), MCF7 (Lane 4), SK-O-V3 (Lane 5) and HEL 92.1.7 (Lane 6) were electrophoresed using NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by XCell SureLock™ Mini-Cell and XCell II™ Blot Module (Product # EI0002). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with 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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Talin 1 was performed using MDA-MB-231 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Talin 1 Goat Polyclonal Antibody (Product # MA5-28133) at 5µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766) at a dilution of 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 composite image showing cytoplasmic and ruffled membrane localization of Talin 1 in MDA-MB-231 cells. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry of Talin in human peripheral blood platelets. Sample was incubated with Talin monoclonal antibody (Product # MA5-28133).
