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RecombinantRabMAb

Recombinant Anti-Galectin 3 antibody [EP2775Y] (ab76245)

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Western blot - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
  • Immunocytochemistry/ Immunofluorescence - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
  • Flow Cytometry - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
  • Western blot - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
  • Western blot - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
  • Immunocytochemistry/ Immunofluorescence - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
  • Flow Cytometry - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
  • 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

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HRP Anti-Galectin 3 antibody [EP2775Y] (ab202553)

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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
    Human
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Mouse
    Human
    See all applications and species data
  • 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
    For more information see here.

    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

    • Neuroscience
    • Cell Adhesion Proteins
    • Nuclear Proteins
    • Cell Biology
    • Cell Cycle
    • Cell differentiation

Associated products

  • Alternative Versions

    • Anti-Galectin 3 antibody [EP2775Y] - BSA and Azide free (ab197544)
    • HRP Anti-Galectin 3 antibody [EP2775Y] (ab202553)
    • Alexa Fluor® 555 Anti-Galectin 3 antibody [EP2775Y] (ab279330)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa nuclear extract lysate (ab14655)
    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
    • A-431 whole cell lysate (ab30132)
    • SW480 whole cell lysate (ab3957)
    • NIH/3T3 whole cell lysate (ab7179)
    • RAW 264.7 whole cell lysate (ab7187)
    • A-431 whole cell lysate (ab7909)
  • Recombinant Protein

    • Recombinant human Galectin 3 protein (Active) (ab50236)

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

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.
  • Target information above from: UniProt accession P17931 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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
    • Unigene: 248615 Mouse
    • Unigene: 764 Rat
    see all
  • Alternative names

    • 35 kDa lectin antibody
    • Carbohydrate binding protein 35 antibody
    • Carbohydrate-binding protein 35 antibody
    • CBP 35 antibody
    • CBP35 antibody
    • Gal-3 antibody
    • GAL3 antibody
    • Galactose-specific lectin 3 antibody
    • Galactoside-binding protein antibody
    • GALBP antibody
    • Galectin 3 internal gene,included antibody
    • Galectin-3 antibody
    • Galectin3 antibody
    • GALIG antibody
    • GBP antibody
    • IgE binding protein antibody
    • IgE-binding protein antibody
    • L 31 antibody
    • L 34 antibody
    • L-31 antibody
    • L-34 galactoside-binding lectin antibody
    • L31 antibody
    • Laminin-binding protein antibody
    • Lectin L-29 antibody
    • Lectin, galactose binding, soluble 3 antibody
    • LEG3_HUMAN antibody
    • LGALS2 antibody
    • LGALS3 antibody
    • MAC 2 antigen antibody
    • Mac-2 antibody
    • Mac-2 antigen antibody
    • MAC2 antibody
    • Macrophage galactose-specific lectin antibody
    • MGC105387 antibody
    see all

Images

  • Western blot - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
    Western blot - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
    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) - 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 - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
    Immunocytochemistry/ Immunofluorescence - Anti-Galectin 3 antibody [EP2775Y] (ab76245)

    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) - 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 - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
    Flow Cytometry - Anti-Galectin 3 antibody [EP2775Y] (ab76245)

    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.

  • Western blot - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
    Western blot - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
    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 kDa



    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
    Western blot - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
    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?

  • Immunocytochemistry/ Immunofluorescence - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
    Immunocytochemistry/ Immunofluorescence - Anti-Galectin 3 antibody [EP2775Y] (ab76245)

    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.

  • Flow Cytometry - Anti-Galectin 3 antibody [EP2775Y] (ab76245)
    Flow Cytometry - Anti-Galectin 3 antibody [EP2775Y] (ab76245)

    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)
    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.

    See Abreview

  • Anti-Galectin 3 antibody [EP2775Y] (ab76245)
    Anti-Galectin 3 antibody [EP2775Y] (ab76245)

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (8)

Publishing research using ab76245? Please let us know so that we can cite the reference in this datasheet.

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

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-5 of 5 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Galectin 3 antibody [EP2775Y]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH6
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde
Read More

Dr. Matthias Dedobbeleer

Verified customer

Submitted Feb 17 2021

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Galectin 3 antibody [EP2775Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Lung
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde
Read More

Mr. Carl Hobbs

Verified customer

Submitted Jul 27 2018

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Galectin 3 antibody [EP2775Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Lung
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde
Read More

Mr. Carl Hobbs

Verified customer

Submitted Jul 27 2018

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Galectin 3 antibody [EP2775Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Lung
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde
Read More

Mr. Carl Hobbs

Verified customer

Submitted Feb 13 2017

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Galectin 3 antibody [EP2775Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Lung
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde
Read More

Mr. Carl Hobbs

Verified customer

Submitted May 17 2016

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