Recombinant Anti-Galectin 3 antibody [EP2775Y] (ab76245)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2775Y] to Galectin 3
- Suitable for: ICC/IF, WB, Flow Cyt, IHC-P
- Reacts with: Mouse, Human
Overview
-
Product name
Anti-Galectin 3 antibody [EP2775Y]
See all Galectin 3 primary antibodies -
Description
Rabbit monoclonal [EP2775Y] to Galectin 3 -
Host species
Rabbit -
Tested Applications & Species
Application Species Flow Cyt HumanICC/IF HumanIHC-P HumanWB MouseHuman -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: A375, HeLa, SW480, A431, RAW264.7 and NIH/3T3 cell lysates. ICC/IF: Panc-1 and HT-29 cells. IHC-P: Human lung squamous carcinoma and human thyroid carcinoma tissues.
-
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP2775Y -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Positive Controls
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab76245 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 |
---|---|
Flow Cyt |
Human
|
ICC/IF |
Human
|
IHC-P |
Human
|
WB |
Mouse
Human
|
All applications |
Rat
Pig
|
Application | Abreviews | Notes |
---|---|---|
ICC/IF |
1/1000.
For unpurified use at 1/100 - 1/250. |
|
WB |
1/5000 - 1/10000. Predicted molecular weight: 26 kDa.
|
|
Flow Cyt |
1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/1000. |
|
IHC-P | (5) |
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/2000 - 1/5000. |
Notes |
---|
ICC/IF
1/1000. For unpurified use at 1/100 - 1/250. |
WB
1/5000 - 1/10000. Predicted molecular weight: 26 kDa. |
Flow Cyt
1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/1000. |
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/2000 - 1/5000. |
Target
-
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
-
Database links
- Entrez Gene: 3958 Human
- Entrez Gene: 16854 Mouse
- Entrez Gene: 83781 Rat
- Omim: 153619 Human
- SwissProt: P17931 Human
- SwissProt: P16110 Mouse
- SwissProt: P08699 Rat
- Unigene: 531081 Human
see all -
Alternative names
- 35 kDa lectin antibody
- Carbohydrate binding protein 35 antibody
- Carbohydrate-binding protein 35 antibody
see all
Images
-
All lanes : Anti-Galectin 3 antibody [EP2775Y] (ab76245) at 1/10000 dilution (purified)
Lane 1 : A375 cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : A431 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 26 kDa
Observed band size: 31 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung squamous carcinoma tissue labelling Galectin 3 with unpurified ab76245.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunocytochemistry/Immunofluorescence analysis of HT-29 cells labelling Galectin with purified ab76245 at 1/1000. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/500) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue labelling Galectin 3 with purified ab76245 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
-
Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Galectin 3 with purified ab76245 at 1/50 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
-
All lanes : Anti-Galectin 3 antibody [EP2775Y] (ab76245) at 1/10000 dilution (purified)
Lane 1 : RAW264.7 cell lysate
Lane 2 : NIH/3T3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 26 kDaBlocking and dilution buffer: 5% NFDM/TBST.
-
All lanes : Anti-Galectin 3 antibody [EP2775Y] (ab76245) at 1/10000 dilution (unpurified)
Lane 1 : A375 cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : SW480 cell lysate
Lane 4 : A431 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 26 kDa
Observed band size: 31 kDa why is the actual band size different from the predicted? -
ICC/IF image of unpurified ab76245 stained Panc-1 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab76245 at 10µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
-
Overlay histogram showing THP1 cells stained with unpurified ab76245 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% human serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76245, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EP2775Y] (ab76245)This image is courtesy of an abreview submitted by Carl Hobbs
Immunohistochemical analysis of formaldehyde fixed mouse lung tissue sections labelling Galectin 3 with ab76245 at a dilution of 1/6000. The secondary antibody used was biotin conjugated goat anti rabbit IgG at a dilution of 1/300. Antigen retrieval was heat mediated using citric acid.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (8)
ab76245 has been referenced in 8 publications.
- Mishra PS et al. Transmission of ALS pathogenesis by the cerebrospinal fluid. Acta Neuropathol Commun 8:65 (2020). PubMed: 32381112
- Sun Z et al. RAGE/galectin-3 yields intraplaque calcification transformation via sortilin. Acta Diabetol N/A:N/A (2019). PubMed: 30603868
- Chen X et al. Increase of glandular epithelial cell clusters by an external volume expansion device promotes adipose tissue regeneration by recruiting macrophages. Biosci Rep 39:N/A (2019). PubMed: 30760630
- Wang H et al. Cathepsin S Deficiency Mitigated Chronic Stress-Related Neointimal Hyperplasia in Mice. J Am Heart Assoc 8:e011994 (2019). PubMed: 31296090
- Ha Y et al. Induction of Lysosome-associated Protein Transmembrane 4 Beta via Sulfatase 2 Enhances Autophagic Flux in Liver Cancer Cells. Hepatol Commun 3:1520-1543 (2019). PubMed: 31701075
- Shi ZR et al. Decrease of galectin-3 in keratinocytes: A potential diagnostic marker and a critical contributor to the pathogenesis of psoriasis. J Autoimmun 89:30-40 (2018). WB, IHC-P ; Mouse, Human . PubMed: 29167025
- Zhao G et al. The Soluble VEGF Receptor sFlt-1 Contributes to Impaired Neovascularization in Aged Mice. Aging Dis 8:287-300 (2017). PubMed: 28580185
- Gao P et al. Anti-inflammatory deficiencies in neutrophilic asthma: reduced galectin-3 and IL-1RA/IL-1ß. Respir Res 16:5 (2015). PubMed: 25616863