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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [1]
- Immunocytochemistry [1]
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- Product number
- 42-5900 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Galectin 1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 0.25 mg/mL
- Storage
- -20°C
Submitted references Pharmacological treatment with galectin-1 protects against renal ischaemia-reperfusion injury.
Carlos CP, Silva AA, Gil CD, Oliani SM
Scientific reports 2018 Jun 22;8(1):9568
Scientific reports 2018 Jun 22;8(1):9568
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Galectin 1 Polyclonal Antibody (Product # 42-5900) and a 15kDa band corresponding to Galectin 1 was observed in MDA-MB-231 and PC-3 but not in LNCaP and MCF7. Whole cell extracts (40 µg lysate) of PC-3 (Lane 1), LNCaP (Lane 2), MCF7 (Lane 3) and MDA-MB-231 (Lane 4) 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 (1:1250 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 cells (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
- 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 # 42-5900) 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 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 and membrane 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.
