Recombinant Anti-HEXB antibody [EPR7978] (ab140649)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7978] to HEXB
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-HEXB antibody [EPR7978]
See all HEXB primary antibodies -
Description
Rabbit monoclonal [EPR7978] to HEXB -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human HEXB aa 400-500. The exact sequence is proprietary.
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Positive control
- WB: Recombinant Human HEXB protein (ab114915) Jurkat, HeLa, HepG2 and Caco-2 cell lysates. IHC-P: Human kidney and lung tissue.
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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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR7978 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab140649 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 |
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WB |
1/1000 - 1/10000. Detects a band of approximately 29, 63 kDa (predicted molecular weight: 63 kDa).
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IHC-P |
1/25 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Antigen retrieval is recommended. |
Notes |
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WB
1/1000 - 1/10000. Detects a band of approximately 29, 63 kDa (predicted molecular weight: 63 kDa). |
IHC-P
1/25 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Antigen retrieval is recommended. |
Target
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Function
Responsible for the degradation of GM2 gangliosides, and a variety of other molecules containing terminal N-acetyl hexosamines, in the brain and other tissues. -
Involvement in disease
Defects in HEXB are the cause of GM2-gangliosidosis type 2 (GM2G2) [MIM:268800]; also known as Sandhoff disease. GM2-gangliosidosis is an autosomal recessive lysosomal storage disease marked by the accumulation of GM2 gangliosides in the neuronal cells. GM2G2 is clinically indistinguishable from GM2-gangliosidosis type 1, presenting startle reactions, early blindness, progressive motor and mental deterioration, macrocephaly and cherry-red spots on the macula. -
Sequence similarities
Belongs to the glycosyl hydrolase 20 family. -
Post-translational
modificationsN-linked glycans at Asn-142 and Asn-190 consist of Man(3)-GlcNAc(2) and Man(5 to 7)-GlcNAc(2), respectively.
The beta-A and beta-B chains are produced by proteolytic processing of the precursor beta chain. -
Cellular localization
Lysosome. - Information by UniProt
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Database links
- Entrez Gene: 3074 Human
- GenBank: NP_000512.1 Human
- Omim: 606873 Human
- SwissProt: P07686 Human
- Unigene: 69293 Human
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Alternative names
- Beta hexosaminidase beta chain antibody
- Beta hexosaminidase subunit beta antibody
- Beta N acetylhexosaminidase antibody
see all
Images
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Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: HEXB knockout HAP1 whole cell lysate (20 µg)
Lane 3: Jurkat whole cell lysate (20 µg)
Lane 4: HeLa whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab140649 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab140649 was shown to specifically react with HEXB when HEXB knockout samples were used. Wild-type and HEXB knockout samples were subjected to SDS-PAGE. Ab140649 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
This image was generated using the unpurified version of the product.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue sections labeling HEXB with Purified ab140649 at 1:500 dilution (4.84 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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All lanes : Anti-HEXB antibody [EPR7978] (ab140649) at 1/1000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 63 kDa
Observed band size: 29,63 kDa why is the actual band size different from the predicted?The double bands caused by proteolytic processing are consistent with what has been described in PMID 2139028
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All lanes : Anti-HEXB antibody [EPR7978] (ab140649) at 1/1000 dilution
Lane 1 : Jurkat cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : Caco-2 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP conjugated goat anti-rabbit at 1/2000 dilution
Predicted band size: 63 kDaThis image was generated using the unpurified version of the product.
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Immunohistochemical analysis of paraffin embedded Human kidney tissue labelling HEXB with ab140649 at a 1/25 dilution.
This image was generated using the unpurified version of the product.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (5)
ab140649 has been referenced in 5 publications.
- Flotte TR et al. AAV gene therapy for Tay-Sachs disease. Nat Med 28:251-259 (2022). PubMed: 35145305
- Adams BM et al. Quantitative glycoproteomics reveals cellular substrate selectivity of the ER protein quality control sensors UGGT1 and UGGT2. Elife 9:N/A (2020). PubMed: 33320095
- Cui Y et al. Retromer has a selective function in cargo sorting via endosome transport carriers. J Cell Biol 218:615-631 (2019). PubMed: 30559172
- Silvers CR et al. Characterization of urinary extracellular vesicle proteins in muscle-invasive bladder cancer. Oncotarget 8:91199-91208 (2017). PubMed: 29207636
- Yasui N et al. Molecular pathology of Sandhoff disease with p.Arg505Gln in HEXB: application of simulation analysis. J Hum Genet 58:611-7 (2013). PubMed: 23759947