Explore
Validate
Learn
Western blot
ELISAAntibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [1]
Submit
Validation data
Reference
Comment
Report error
- Product number
- MA5-15295 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- LCK Monoclonal Antibody (8E5F9, 4B2C7)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- MA5-15295 targets Lck in indirect ELISA, WB applications and shows reactivity with Human samples. The MA5-15295 immunogen is purified recombinant fragment of human Lck expressed in E. Coli. MA5-15295 detects Lck which has a predicted molecular weight of approximately 56kDa.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 8E5F9, 4B2C7
- Vial size
- 100 µL
- Concentration
- Conc. Not Determined
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Lck using Lck monoclonal antibody (Product # MA5-15295) in MOLT-4 (1), CCRF-CEM (2), CCRF-HSB-2 (3) and Jurkat (4) cell lysate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Lck using Lck monoclonal antibody (Product # MA5-15295) in MOLT-4 (1), CCRF-CEM (2), CCRF-HSB-2 (3) and Jurkat (4) cell lysate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using LCK Monoclonal Antibody (8E5F9, 4B2C7) (Product # MA5-15295) and a 55 kDa band corresponding to LCK was observed in positive cell lines Jurkat, Ramos and tissues such as Mouse Thymus, Mouse Spleen and not in negative cell lines such as A549 and Mouse brain as mentioned in literature. Membrane enriched extracts (30 µg lysate) of Jurkat (Lane 1), Ramos (Lane 2), A549 (Lane 3), Mouse Thymus (Lane 4), Mouse Brain (Lane 5) and Mouse Spleen (Lane 6) were electrophoresed using NuPAGE® 4-12 % Bis-Tris 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-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 LCK was performed using Jurkat 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 LCK Monoclonal Antibody (8E5F9, 4B2C7) (Product # MA5-15295) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) 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 merged image showing membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
