Recombinant Anti-Galectin 3 antibody [EPR19244] - BSA and Azide free (ab251504)
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 - BSA and Azide free
- Suitable for: Flow Cyt, WB, IP, IHC-P, ICC
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-Galectin 3 antibody [EPR19244] - BSA and Azide free
See all Galectin 3 primary antibodies -
Description
Rabbit monoclonal [EPR19244] to Galectin 3 - BSA and Azide free -
Host species
Rabbit -
Tested Applications & Species
Application Species Flow Cyt HumanIHC-P HumanIP HumanWB Human -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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General notes
Ab251504 is the carrier-free version of ab209344. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.
Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
ab251504 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.
Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
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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Conjugation kits
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab251504 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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Flow Cyt |
Human
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IHC-P |
Human
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IP |
Human
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WB |
Human
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Application | Abreviews | Notes |
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Flow Cyt |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa).
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa). |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC
Use at an assay dependent concentration. |
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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This data was developed using ab209344, the same antibody clone in a different buffer formulation.
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: alectin 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.
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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 kDaThis data was developed using ab209344, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times: Lanes 1-2: 30 seconds; Lanes 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 kDaThis data was developed using ab209344, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times: Lanes 1-2: 15 seconds; Lane 3: 30 seconds.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EPR19244] - BSA and Azide free (ab251504)
This data was developed using ab209344, the same antibody clone in a different buffer formulation.
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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EPR19244] - BSA and Azide free (ab251504)
This data was developed using ab209344, the same antibody clone in a different buffer formulation.
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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EPR19244] - BSA and Azide free (ab251504)
This data was developed using ab209344, the same antibody clone in a different buffer formulation.
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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EPR19244] - BSA and Azide free (ab251504)This data was developed using ab209344, the same antibody clone in a different buffer formulation.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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This data was developed using ab209344, the same antibody clone in a different buffer formulation.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. -
This data was developed using ab209344, the same antibody clone in a different buffer formulation.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. -
This data was developed using ab209344, the same antibody clone in a different buffer formulation.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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This data was developed using ab209344, the same antibody clone in a different buffer formulation.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
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab251504 has not yet been referenced specifically in any publications.