Anti-galectin 9/Gal-9 antibody [EPR22214] (ab227046)
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
- Suitable for: ICC, WB, IHC-P, Flow Cyt
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-galectin 9/Gal-9 antibody [EPR22214]
See all galectin 9/Gal-9 primary antibodies -
Description
Rabbit monoclonal [EPR22214] to galectin 9/Gal-9 -
Host species
Rabbit -
Specificity
This antibody is not recommended for mouse and rat in IHC.
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Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC HumanIHC-P HumanWB 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. WB: U937, HL-60 and THP-1 whole cell lysates. ICC/IF: U937 cells. Flow Cyt: U937 cells.
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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.
Properties
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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: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22214 -
Isotype
IgG -
Research areas
Images
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Immunocytochemistry analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Cell line 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).
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).Confocal image showing nuclear and cytoplasmic staining in U937 cell line.
Negative control: PANC-1.
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All lanes : Anti-galectin 9/Gal-9 antibody [EPR22214] (ab227046) at 1/500 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : Galectin 9 knockout A549 cell lysate
Lane 3 : U-937 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 40 kDa
Observed band size: 34-39 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab227046 observed at 34-39 kDa. Red - loading control ab8245 observed at 36 kDa.
ab227046 Anti-galectin 9/Gal-9 antibody [EPR22214] was shown to specifically react with galectin 9/Gal-9 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266923 (knockout cell lysate ab256976) was used. Wild-type and galectin 9/Gal-9 knockout samples were subjected to SDS-PAGE. ab227046 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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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). -
All lanes : Anti-galectin 9/Gal-9 antibody [EPR22214] (ab227046) at 1/1000 dilution
Lane 1 : U937 (human histiocytic lymphoma monocyte), whole cell lysate
Lane 2 : HL-60 (human Acute Promyelocytic Leukemia promyeloblast), whole cell lysate
Lane 3 : MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate
Lane 4 : PANC-1 (human pancreatic epithelioid carcinoma epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 40 kDa
Observed band size: 34-39 kDa why is the actual band size different from the predicted?
Additional bands at: 13-15 kDa (possible degradation product)
Exposure time: 59 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The expression of the isoforms observed is consistent with what has been described in the literature (PMID 11886844, PMID: 22805533).
Degraded fragments (13-15 kDa) have also been descried (PMID: 15811318).
Negative control: PANC-1.
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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.
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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). -
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. -
All lanes : Anti-galectin 9/Gal-9 antibody [EPR22214] (ab227046) at 1/1000 dilution
Lane 1 : THP-1 (human monocytic leukemia monocyte), whole cell lysate
Lane 2 : HEK-293T (human embryonic kidney epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 40 kDa
Observed band size: 34-39 kDa why is the actual band size different from the predicted?
Additional bands at: 13-15 kDa (possible degradation product)
Exposure time: 59 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The expression of the isoforms observed is consistent with what has been described in the literature (PMID 11886844, PMID: 22805533).
Degraded fragments (13-15 kDa) has also been described (PMID: 15811318).
Negative control: HEK-293T (PMID: 24333696).
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