Name: Anti-Galectin 3 antibody [EPR19244]
Brand: RabMAb
SKU: ab209344
Rating: 5 (5 reviews)

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Western blot - 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

Carrier Free

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.
Positive control
- WB: HeLa, SW480, MCF7 and A431 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.
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.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19244
Isotype

IgG
Research areas

Alternative Versions
- Anti-Galectin 3 antibody [EPR19244] - BSA and Azide free (ab251504)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (ab210849)
Recombinant Protein
- Recombinant human Galectin 3 protein (Active) (ab50236)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

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
Flow Cyt (Intra)		1/50.
IP		1/30.
ICC/IF	(3)	1/500.
IHC-P	(1)	1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB	(1)	1/1000. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa).

Notes
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).

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
- Omim: 153619 Human
- SwissProt: P17931 Human
- Unigene: 531081 Human
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

Western blot - Anti-Galectin 3 antibody [EPR19244] (ab209344)

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.
Western blot - Anti-Galectin 3 antibody [EPR19244] (ab209344)

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 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1/2: 30 seconds; Lane 3/4: 8 seconds
Western blot - Anti-Galectin 3 antibody [EPR19244] (ab209344)

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 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1/2: 15 seconds; Lane 3: 30 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EPR19244] (ab209344)

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] (ab209344)

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] (ab209344)

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] (ab209344)

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.
Immunocytochemistry/ Immunofluorescence - Anti-Galectin 3 antibody [EPR19244] (ab209344)

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.
Immunocytochemistry/ Immunofluorescence - Anti-Galectin 3 antibody [EPR19244] (ab209344)

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.
Flow Cytometry (Intracellular) - Anti-Galectin 3 antibody [EPR19244] (ab209344)

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.
Immunoprecipitation - Anti-Galectin 3 antibody [EPR19244] (ab209344)

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.
Anti-Galectin 3 antibody [EPR19244] (ab209344)

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

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab209344? Please let us know so that we can cite the reference in this datasheet.

ab209344 has been referenced in 1 publication.

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

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1-5 of 5 Abreviews or Q&A

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (PC-3 cells - human prostate cancer cells)

Permeabilization

Yes - 0.2% Triton X-100 in PBS

Specification

PC-3 cells - human prostate cancer cells

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C

Fixative

Formaldehyde

Dr. Armen Petrosyan

Verified customer

Submitted Apr 08 2022

Application

Immunocytochemistry/ Immunofluorescence

Sample

Rat Cell (Rat hepatocytes)

Permeabilization

Yes - 0.2% Triton X-100 in PBS

Specification

Rat hepatocytes

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C

Fixative

Formaldehyde

Dr. Armen Petrosyan

Verified customer

Submitted Apr 06 2022

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Human prostate tissue)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris-EDTA buffer (pH 9.0)

Permeabilization

Yes - 0.2% Triton X-100 in PBS

Specification

Human prostate tissue

Blocking step

(agent) for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 37°C

Fixative

Formaldehyde

Dr. Armen Petrosyan

Verified customer

Submitted Nov 29 2021

Application

Immunocytochemistry/ Immunofluorescence

Sample

Mouse Cell (Mice hepatocytes)

Permeabilization

Yes - 0.2% Triton X-100 in PBS

Specification

Mice hepatocytes

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C

Fixative

Formaldehyde

Dr. Armen Petrosyan

Verified customer

Submitted Oct 07 2021

Application

Western blot

Sample

Human Cell lysate - other (PC-3 cells - human prostate cancer cells - plasma)

Gel Running Conditions

Reduced Denaturing (4-15% SDS-PAGE)

Loading amount

40 µg

Specification

PC-3 cells - human prostate cancer cells - plasma

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Dr. Armen Petrosyan

Verified customer

Submitted Aug 11 2021

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (1)

Customer reviews and Q&As

Immunocytochemistry/ Immunofluorescence abreview for Anti-Galectin 3 antibody [EPR19244]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Galectin 3 antibody [EPR19244]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Galectin 3 antibody [EPR19244]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Galectin 3 antibody [EPR19244]

Western blot abreview for Anti-Galectin 3 antibody [EPR19244]