Recombinant Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)
![Western blot - Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)](https://www.abcam.com/ps/products/243/ab243288/Images/ab243288-484795-ab227046464321antigalectin9gal9antibodyepr22214westernblotwildtypethp1lgals9knockoutthp1.jpg "Western blot - Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22214] to galectin 9/Gal-9 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), IHC-P, WB, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free
See all galectin 9/Gal-9 primary antibodies -
Description
Rabbit monoclonal [EPR22214] to galectin 9/Gal-9 - BSA and Azide free -
Host species
Rabbit -
Specificity
This antibody is not recommended for mouse and rat in IHC.
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Tested applications
Suitable for: Flow Cyt (Intra), IHC-P, WB, ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human liver and colon tissue. ICC/IF: U937 cells. Flow Cyt (intra): U937 cells. WB: THP-1 whole cell lysates
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General notes
ab243288 is the carrier-free version of ab227046.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
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.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22214 -
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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Immunohistochemistry reagents
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab243288 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) |
Use at an assay dependent concentration.
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| IHC-P | (1) |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
This antibody is not recommended for human in IHC. |
| WB |
Use at an assay dependent concentration. Predicted molecular weight: 40 kDa.
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| ICC/IF |
Use at an assay dependent concentration.
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| Notes |
|---|
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Flow Cyt (Intra)
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. This antibody is not recommended for human in IHC. |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 40 kDa. |
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ICC/IF
Use at an assay dependent concentration. |
Target
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Function
Binds galactosides. Has high affinity for the Forssman pentasaccharide. May play a role in thymocyte-epithelial interactions relevant to the biology of the thymus. Inhibits cell proliferation. It is a ligand for HAVCR2/TIM3. Induces T-helper type 1 lymphocyte (Th1) death. Isoform Short acts as an eosinophil chemoattractant. -
Tissue specificity
Peripheral blood leukocytes and lymphatic tissues. Overexpressed in Hodgkin disease tissue. -
Sequence similarities
Contains 2 galectin domains. -
Domain
Contains two homologous but distinct carbohydrate-binding domains. -
Cellular localization
Cytoplasm. Secreted. May also be secreted by a non-classical secretory pathway. - Information by UniProt
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Database links
- Entrez Gene: 3965 Human
- Omim: 601879 Human
- SwissProt: O00182 Human
- Unigene: 81337 Human
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Alternative names
- 36 kDa beta-galactoside-binding lectin antibody
- Ecalectin antibody
- Gal-9 antibody
see all
Images
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Western blot - Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)All lanes : Anti-galectin 9/Gal-9 antibody [EPR22214] (ab227046) at 1/1000 dilution
Lane 1 : Wild-type THP-1 cell lysate
Lane 2 : LGALS9 knockout THP-1 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 40 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab227046 observed at 35 kDa. Red - loading control Mouse anti Histone H3 observed at 18 kDa.
ab227046 was shown to react with galectin 9/Gal-9 in wild-type THP-1 cells in Western blot with loss of signal observed in LGALS9 knockout cell line ab269505 (knockout cell lysate ab269667). Wild-type THP-1 and LGALS9 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab227046 and Mouse anti Histone H3 overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)](https://www.abcam.com/ps/products/243/ab243288/Images/ab243288-332653-anti-galectin-9-gal-9-antibody-epr22214-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)Immunohistochemical analysis of paraffin-embedded human liver tissue labeling galectin 9/Gal-9 with ab227046 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear and cytoplasmic staining on Kupffer cells of human liver (PMID: 20209097, PMID: 18202194) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227046).
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![Immunocytochemistry/ Immunofluorescence - Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)](https://www.abcam.com/ps/products/243/ab243288/Images/ab243288-332650-anti-galectin-9-gal-9-antibody-epr22214-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U937 (human histiocytic lymphoma cell line) cells labeling galectin 9/Gal-9 with ab227046 at 1/250 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 in U-937 cells. The nuclear counterstain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
PANC-1 (human pancreatic epithelial cancinoma cell line) cells stained in the same manner serve as a negative control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227046).
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![Flow Cytometry (Intracellular) - Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)](https://www.abcam.com/ps/products/243/ab243288/Images/ab243288-332652-anti-galectin-9-gal-9-antibody-epr22214-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized PANC-1 (human pancreatic epithelial cancinoma cell line) (Left) / U937 (human histiocytic lymphoma cell line) (Right) cell line labeling galectin 9/Gal-9 with ab227046 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor®488, ab150077), at 1/2000 dilution was used as the secondary antibody.
Negative Control -PANC-1 cells.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227046).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)](https://www.abcam.com/ps/products/243/ab243288/Images/ab243288-332654-anti-galectin-9-gal-9-antibody-epr22214-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)Immunohistochemical analysis of paraffin-embedded human colon tissue labeling galectin 9/Gal-9 with ab227046 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear and cytoplasmic staining on epithelial cells of human colon (PMID: 18202194) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227046).
-
![Immunocytochemistry/ Immunofluorescence - Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)](https://www.abcam.com/ps/products/243/ab243288/Images/ab243288-332651-anti-galectin-9-gal-9-antibody-epr22214-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney) cells labeling galectin 9/Gal-9 with ab227046 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing no staining in HEK-293T cells
Negative control: HEK-293T (PMID: 11698107). The nuclear counterstain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of the primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227046).
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![Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)](https://www.abcam.com/ps/products/243/ab243288/Images/ab243288-7-benefits-of-recombinant-antibodies.png)
Anti-galectin 9/Gal-9 antibody [EPR22214] - BSA and Azide free (ab243288)
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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Datasheet download
Certificate of Compliance
References (0)
ab243288 has not yet been referenced specifically in any publications.
