Recombinant Anti-Galectin 3 antibody [EPR19244] (ab209344)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19244] to Galectin 3
- Suitable for: Flow Cyt (Intra), IP, ICC/IF, IHC-P, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Galectin 3 antibody [EPR19244]
See all Galectin 3 primary antibodies -
Description
Rabbit monoclonal [EPR19244] to Galectin 3 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IP, ICC/IF, IHC-P, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, SW480, MCF7, A431 and Wild-type A549 whole cell lysates; human heart, kidney and stomach lysates. IHC-P: Human liver, stomach, diffuse large B cell lymphoma and colon cancer tissues. ICC/IF: HeLa and A431 cells. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19244 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab209344 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/50.
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IP |
1/30.
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ICC/IF | (3) |
1/500.
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IHC-P | (1) |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB | (1) |
1/1000. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa).
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Notes |
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Flow Cyt (Intra)
1/50. |
IP
1/30. |
ICC/IF
1/500. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa). |
Target
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Function
Galactose-specific lectin which binds IgE. May mediate with the alpha-3, beta-1 integrin the stimulation by CSPG4 of endothelial cells migration. Together with DMBT1, required for terminal differentiation of columnar epithelial cells during early embryogenesis. -
Tissue specificity
A major expression is found in the colonic epithelium. It is also abundant in the activated macrophages. -
Sequence similarities
Contains 1 galectin domain. -
Cellular localization
Nucleus. Cytoplasmic in adenomas and carcinomas. May be secreted by a non-classical secretory pathway and associate with the cell surface. - Information by UniProt
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Database links
- Entrez Gene: 3958 Human
- Omim: 153619 Human
- SwissProt: P17931 Human
- Unigene: 531081 Human
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Alternative names
- 35 kDa lectin antibody
- Carbohydrate binding protein 35 antibody
- Carbohydrate-binding protein 35 antibody
see all
Images
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All lanes : Anti-Galectin 3 antibody [EPR19244] (ab209344) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : LGALS3 knockout A549 cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : LNCaP cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 26 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Galectin 3 antibody [EPR19244] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab209344 was shown to bind specifically to Galectin 3. A band was observed at 30 kDa in wild-type A549 cell lysates with no signal observed at this size in LGALS3 knockout cell line. To generate this image, wild-type and LGALS3 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Galectin 3 (KO) knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: MCF7 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab209344 observed at 35 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab209344 was shown to specifically react with Galectin 3 (KO) in wild-type HAP1 cells. No band was observed when LGALS3 (KO) knockout samples were examined. Wild-type and Galectin 3 (KO) knockout samples were subjected to SDS-PAGE. Ab209344 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging. -
Lanes 1-2 : Anti-Galectin 3 antibody [EPR19244] (ab209344) at 1/10000 dilution
Lanes 3-4 : Anti-Galectin 3 antibody [EPR19244] (ab209344) at 1/1000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 4 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 26 kDa
Observed band size: 26 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1/2: 30 seconds; Lane 3/4: 8 seconds
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All lanes : Anti-Galectin 3 antibody [EPR19244] (ab209344) at 1/1000 dilution
Lane 1 : Human heart tissue
Lane 2 : Human kidney tissue
Lane 3 : Human stomach tissue
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 26 kDa
Observed band size: 26 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1/2: 15 seconds; Lane 3: 30 seconds
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Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Galectin 3 with ab209344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Positive staining on Kupffer cells in the liver is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling Galectin 3 with ab209344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and nuclear staining on human stomach is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded human diffuse large B cell lymphoma tissue labeling Galectin 3 with ab209344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and weak nuclear staining on human diffuse large B cell lymphoma is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling Galectin 3 with ab209344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and weak nuclear staining on human colon cancer is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Galectin 3 with ab209344 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L(Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weak cytoplasmic staining on HeLa cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab209344 at 1/500 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L(Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma cell line) cells labeling Galectin 3 with ab209344 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weakly cytoplasmic staining on A431 cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab209344 at 1/500 dilution followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. -
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Galectin 3 with ab209344 at 1/50 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
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Galectin 3 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab209344 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab209344 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HeLa whole cell lysate 10µg (Input).
Lane 2: ab209344 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab209344 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (5)
ab209344 has been referenced in 5 publications.
- Rahrmann EP et al. The NALCN channel regulates metastasis and nonmalignant cell dissemination. Nat Genet 54:1827-1838 (2022). PubMed: 36175792
- Chen X et al. Intracellular galectin-3 is a lipopolysaccharide sensor that promotes glycolysis through mTORC1 activation. Nat Commun 13:7578 (2022). PubMed: 36481721
- Cano I et al. Galectin-3 Enhances Vascular Endothelial Growth Factor-A Receptor 2 Activity in the Presence of Vascular Endothelial Growth Factor. Front Cell Dev Biol 9:734346 (2021). PubMed: 34616740
- Lu L et al. Angiotensin (ang) 1-7 inhibits ang II-induced atrial fibrosis through regulating the interaction of proto-oncogene tyrosine-protein kinase Src (c-Src) and Src homology region 2 domain-containing phosphatase-1 (SHP-1)). Bioengineered 12:10823-10836 (2021). PubMed: 34872449
- Li Y et al. Integrated Bioinformatics Analysis Reveals Key Candidate Genes and Pathways Associated With Clinical Outcome in Hepatocellular Carcinoma. Front Genet 11:814 (2020). PubMed: 32849813