Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [5]
- Immunocytochemistry [4]
- Immunohistochemistry [2]
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Validation data
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- Product number
- PA5-27455 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Galectin 1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: K562, THP-1, HeLa, NCI-H929, BCL-1, PC-12, Rat2, U2OS. Predicted reactivity: Mouse (86%), Rat (89%), Dog (81%), Pig (84%), Sheep (84%), Rhesus Monkey (97%), Bovine (85%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.52 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- 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 # PA5-27455) at a dilution of 1:10000 and a HRP-conjugated anti-rabbit IgG secondary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using Galectin 1 Polyclonal Antibody (Product # PA5-27455). Sample (30 µg of whole cell lysate). Lane A: NCI-H929. 15% SDS PAGE. Galectin 1 Polyclonal Antibody (Product # PA5-27455) diluted at 1:10,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using Galectin 1 Polyclonal Antibody (Product # PA5-27455). Sample (30 µg of whole cell lysate). Lane A: BCL-1. 15% SDS PAGE. Galectin 1 Polyclonal Antibody (Product # PA5-27455) diluted at 1:10,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- 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 (25ug 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 # PA5-27455, 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Galectin 1 Polyclonal Antibody (Product # PA5-27455) and a 15kDa band corresponding to Galectin 1 was observed in MDA-MB-231, THP-1 and PC-3 but not in MCF7 and LNCaP. Whole cell extracts (30 µg lysate) of MDA-MB-231 (Lane 1), MCF7 (Lane 2), THP-1 (Lane 3), LNCaP (Lane 4) and PC-3 (Lane 5) 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 # A27036,1:8000 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)
- Main image
- Experimental details
- Immunofluorescent analysis of Galectin 1 showing staining in the cytoplasm of A549 cells. A549 cells were fixed in 4% paraformaldehyde at RT for 15 min and stained using a Galectin 1 polyclonal antibody (Product # PA5-27455) diluted at 1:500. Blue: Hoechst 33342 staining.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry-Immunofluorescence analysis of Galectin 1 was performed in SK N SH cells fixed in 4% paraformaldehyde at RT for 15 min. Green: Galectin 1 Polyclonal Antibody (Product # PA5 27455) diluted at 1:1000. Blue: Hoechst 33342 staining. Scale bar = 10 µm.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Galectin 1 Polyclonal Antibody detects Galectin 1 protein at cytoplasm by immunofluorescent analysis. Sample: SK-N-SH cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: Galectin 1 stained by Galectin 1 Polyclonal Antibody (Product # PA5-27455) diluted at 1:1,000. Blue: Fluoroshield with DAPI . Scale bar= 10 µm.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Galectin 1 was performed using 70% confluent log phase MDA-MB-231 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 # PA5-27455) at 1:100 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), 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 and membrane structures localization. Panel e represents MCF7 cells showing negative staining for Galectin-1. 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
- Immunohistochemistry (Frozen) analysis of Galectin-1 was performed in frozen sectioned E13.5 Rat brain tissue using Galectin 1 Polyclonal Antibody (Product # PA5-27455) at a dilution of 1:250 (Green). Red: beta Tubulin 3/ TUJ1, a mature neuron marker, stained by beta Tubulin 3/ TUJ1 antibody diluted at 1:500. Blue: Fluoroshield with DAPI.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Galectin 1 (Product # PA5-27455) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.