- PA5-30706 - Invitrogen Antibodies LGALS1 antibody

Meade ML, Hoffmann A, Makley MK, Snider TH, Schlager JJ, Gearhart JM
Brain research 2015 Jun 22;1611:101-13

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of Galectin 1 was performed by separating 30 µg of various whole cell extracts by 15% SDS-PAGE. Proteins were transferred to a membrane and probed with a Galectin 1 Polyclonal Antibody (Product # PA5-30706) at a dilution of 1:2000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Galectin 1 Polyclonal Antibody (Product # PA5-30706). Sample (30 µg of whole cell lysate). Lane A: NCI-H929.15% SDS PAGE. Galectin 1 Polyclonal Antibody (Product # PA5-30706) diluted at 1:10,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Galectin 1 Polyclonal Antibody detects Galectin 1 protein by western blot analysis. A. 30 µg PC-12 whole cell lysate/extract. B. 30 µg Rat2 whole cell lysate/extract.15% SDS-PAGE. Galectin 1 Polyclonal Antibody (Product # PA5-30706) dilution: 1:10,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Galectin 1 Polyclonal Antibody (Product # PA5-30706). Sample (30 µg of whole cell lysate). Lane A: BCL-1. 15% SDS PAGE. Galectin 1 Polyclonal Antibody (Product # PA5-30706) diluted at 1:10,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of Galectin 1 was achieved by transfecting PC-3 with Galectin 1 specific siRNAs (Silencer® select Product # s194592, s8145). Western blot analysis (Fig. a) was performed using whole cell extracts (25 ug lysate) from the Galectin 1 knockdown cells (Lane 3), non-targeting scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Galectin 1 Polyclonal Antibody (Product # PA5-30706, 1:25000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:8000 dilution). 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 1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-Galectin 1 Polyclonal Antibody (Product # PA5-30706) and a 15kDa band corresponding to Galectin 1 was observed in MDA-MB-231, THP-1, PC-3 and Caki-1 but not in MCF7, LNCaP. Whole cell extracts (30 µg lysate) of MDA-MB-231 (Lane 1), MCF7 (Lane 2), THP-1 (Lane 3), LNCaP (Lane 4), PC-3 (Lane 5) and Caki-1 (Lane 6) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). 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 (1:15000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Increased expression of Galectin-1 is seen in triple negative breast cancer (TNCB) (like MDA-MB-231) for promoting metastasis but not in non-TNBC like MCF7 [10.18632/oncotarget.16208] and in androgen unresponsive PC-3 cells in comparison to androgen responsive LNCaP [10.1155/2013/519436].

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of Galectin 1 in methanol-fixed A431 cells using a Galectin 1 polyclonal antibody (Product # PA5-30706) at a 1:200 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of Galectin 1 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: Galectin 1 Polyclonal Antibody (Product # PA5 30706) diluted at 1:200. Red: phalloidin, a cytoskeleton marker. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of Galectin 1 was performed in A549 cells fixed in 4% paraformaldehyde at RT for 15 min. Green: Galectin 1 Polyclonal Antibody (Product # PA5 30706) diluted at 1:500. Red: phalloidin, a cytoskeleton marker. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Galectin 1 was performed using 70% confluent log phase MDA-MB-231 and MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Galectin 1 Polyclonal Antibody (Product # PA5-30706) at 1:200 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2500 dilution), 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 cytoplasmic localization of Galectin 1 in MDA-MB-231. Panel e represents MCF7 cells showing negative staining for the same. Panel f represents control MDA-MB-231 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: Galectin 1 Polyclonal Antibody immunoprecipitates Galectin 1 protein in IP experiments. IP Sample: HeLa whole cell lysate/extract. A : 30 µg whole cell lysate/extract of Galectin 1 Polyclonal Antibody expressing HeLa cells. B : Control with 2.5 µg of pre-immune rabbit IgG. C : Immunoprecipitation of Galectin 1 by 2.5 µg of Galectin 1 Polyclonal Antibody (Product # PA5-30706). 15% SDS-PAGE. The immunoprecipitated Galectin 1 protein was detected by Galectin 1 Polyclonal Antibody (Product # PA5-30706) diluted at 1:1,000. Anti-rabbit IgG (HRP) was used as a secondary reagent.

PA5-30706