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Western blot
ELISAAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Immunohistochemistry [3]
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- Product number
- MA1-19344 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Blood Group Lewis A Monoclonal Antibody (7LE)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- For IHC (P), heat mediated antigen retrieval in sodium citrate is recommended.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 7LE
- Vial size
- 100 µg
- Concentration
- 1 mg/mL
- Storage
- 4° C, do not freeze
Submitted references High-throughput carbohydrate microarray profiling of 27 antibodies demonstrates widespread specificity problems.
Manimala JC, Roach TA, Li Z, Gildersleeve JC
Glycobiology 2007 Aug;17(8):17C-23C
Glycobiology 2007 Aug;17(8):17C-23C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Blood Group Lewis A Monoclonal Antibody (Product # MA1-19344) and bands spanning between 60-200 kDa corresponding to glycosylated forms of Blood Group Lewis A were observed in A-431 and were absent in SK-BR-3, MCF-7 and PC-3 cell lysates. Whole cell extracts (30 µg lysate) of A-431 (Lane 1), SK-BR-3 (Lane 2), MCF-7 (Lane 3) and PC-3 (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 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 Blood Group Lewis A was performed using 70% confluent log phase A-431 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 Blood Group Lewis A Monoclonal Antibody (7LE) (Product # MA1-19344) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant 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 SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing plasma membrane and cytoplasmic localization. Panel e shows MCF-7 cells with no expression of Blood Group Lewis A. Panel f 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
- Immunohistochemical analysis of Blood Group Lewis A using a monoclonal antibody (Product # MA1-19344).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry staining of human colon adenocarcinoma (paraffin-embedded sections) with anti-Blood Group Lewis a (7LE) Monoclonal antibody (Product # MA1-19344).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry staining of human pancreas (paraffin-embedded sections) with anti-Blood Group Lewis a (7LE) Monoclonal antibody (Product # MA1-19344).
