MA1-40229

antibody from Invitrogen Antibodies

Targeting: LGALS3 GALIG, LGALS2, MAC-2

ELISA

Immunohistochemistry

Flow cytometry

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Antibody data

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Validation data

Product number: MA1-40229 - Provider product page
Provider: Invitrogen Antibodies
Product name: Galectin 3 Monoclonal Antibody (B2C10)
Antibody type: Monoclonal
Antigen: Other
Description: MA1-40229 detects Galectin-3 from human and mouse samples. This antibody does not cross react with Galedin-3C and is cross reactive with mouse galectin-3. MA1-40229 has been successfully used in flow cytometry, immunohistochemistry (frozen), immunohistochemistry (paraffin), functional assays, ELISA and Western blot applications. The MA1-40229 immunogen is the first 45 residues of human Galectin-3. Endotoxin
Reactivity: Human, Mouse
Host: Mouse
Isotype: IgG
Antibody clone number: B2C10
Vial size: 100 µg
Concentration: 0.1 mg/mL
Storage: 4° C

LGALS3 protein structure - MA1-40229 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of Galectin3 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR855199_LV and CRISPR855204_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of Galectin3 was performed by loading 30 µg of HeLa Wild Type (Lane 1), HeLa Cas9 (Lane 2) andHela Galectin3 KO (Lane 3) membrane enriched extracts. The samples were electrophoresed using NuPAGE™ Novex™ 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 Anti-Galectin 3 Monoclonal Antibody (B2C10) (Product # MA1-40229, 1:200 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:6,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to Galectin3.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Galectin 3 Monoclonal Antibody (B2C10)(Product # MA1-40229) and a ~30kDa band corresponding to Galectin 3 was observed across cell lines tested . Whole cell extracts (30 µg lysate) of HeLa (Lane 1), MDA-MB-231 (Lane 2), A549 (Lane 3), PC-3 (Lane 4), U-87 MG (Lane 5), A-431 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:200) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000) 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: Knockdown of Galectin 3 was achieved by transfecting MDA-MB-231 cells with Galectin 3 specific siRNAs (Silencer® select Product # S8149, S8148). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Galectin 3 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Galectin 3 Monoclonal Antibody (B2C10) (Product # MA1-40229, 1:100 ) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Galectin 3.