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AB324803

Anti-ASAH1 antibody [MJF-D29767-517] - BSA and Azide free

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Rabbit Recombinant Monoclonal ASAH1 antibody. Carrier free. Suitable for Flow Cyt (Intra), ICC/IF, IP and reacts with Human samples.

View Alternative Names

ASAH, HSD-33, HSD33, ASAH1, Acid ceramidase, AC, ACDase, Acid CDase, Acylsphingosine deacylase, Glycosylceramide deacylase, N-acylethanolamine hydrolase ASAH1, N-acylsphingosine amidohydrolase, Putative 32 kDa heart protein, PHP32

4 Images
Immunocytochemistry/ Immunofluorescence - Anti-ASAH1 antibody [MJF-D29767-517] - BSA and Azide free (AB324803)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ASAH1 antibody [MJF-D29767-517] - BSA and Azide free (AB324803)

This data was developed using ab322109, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed 0.1% TritonX-100 permeabilized SK-MEL-28 (human malignant melanoma cell) cells labelling ASAH1 with ab322109 at 1/50 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in SK-MEL-28 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Negative control : A375 (PMID : 26553872). Image was taken with a confocal microscope (Leica-Microsystems TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

Immunocytochemistry/ Immunofluorescence - Anti-ASAH1 antibody [MJF-D29767-517] - BSA and Azide free (AB324803)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ASAH1 antibody [MJF-D29767-517] - BSA and Azide free (AB324803)

This data was developed using ab322109, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed 0.1% TritonX-100 permeabilized MeWo(human malignant melanoma fibroblast) cells labelling ASAH1 with ab322109 at 1/50 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in MeWo cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Negative control : NCI-H1299 (PMID : 26553872). Image was taken with a confocal microscope (Leica-Microsystems TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

Flow Cytometry (Intracellular) - Anti-ASAH1 antibody [MJF-D29767-517] - BSA and Azide free (AB324803)
  • Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-ASAH1 antibody [MJF-D29767-517] - BSA and Azide free (AB324803)

This data was developed using ab322109, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A375 (human malignant melanoma epithelial cell Left) / SK-MEL-28 (human malignant melanoma cell Right) cells labelling ASAH1 with ab322109 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody). Goat Anti-Rabbit IgG (Alexa Fluor® 488 ab150081) at 1/5000 dilution was used as the secondary antibody. Negative control : A375 (PMID : 26553872).

Immunoprecipitation - Anti-ASAH1 antibody [MJF-D29767-517] - BSA and Azide free (AB324803)
  • IP

Supplier Data

Immunoprecipitation - Anti-ASAH1 antibody [MJF-D29767-517] - BSA and Azide free (AB324803)

This data was developed using ab322109, the same antibody clone in a different buffer formulation.

ASAH1 was immunoprecipitated from 0.35 mg SK-MEL-28 (human malignant melanoma cell ) whole cell lysate with ab322109 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322109 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : SK-MEL-28 (human malignant melanoma cell ) whole cell lysate
Lane 2 : ab322109 IP in SK-MEL-28 (human malignant melanoma cell ) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab322109 in SK-MEL-28 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST

The IP experiment was performed by ab322109 using SK-MEL-28 lysate. The blot was probed by another anti-ASAH1 antibody (ab282276)(1 : 1000 dilution).

All lanes:

Immunoprecipitation - Anti-ASAH1 antibody [MJF-D29767-517] (<a href='/en-us/products/primary-antibodies/asah1-antibody-mjf-d29767-517-ab322109'>ab322109</a>) at 1/30 dilution

All lanes:

SK-MEL-28 (human malignant melanoma cell ) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

MJF-D29767-517

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

Flow Cyt (Intra), IP, ICC/IF

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCFr" : {"fullname" : "Immunohistochemistry (Frozen sections)", "shortname":"IHC-Fr"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p></p>", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" }, "Mouse": { "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "<p></p>" }, "Rat": { "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "<p></p>" } } }
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Product details

ab324803 is the carrier-free version of ab322109.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

Compatibility
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.

Collaborations
This antibody was developed with support from The Michael J. Fox Foundation.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Lysosomal ceramidase that hydrolyzes sphingolipid ceramides into sphingosine and free fatty acids at acidic pH (PubMed : 10610716, PubMed : 11451951, PubMed : 15655246, PubMed : 26898341, PubMed : 36752535, PubMed : 7744740, PubMed : 7852294). Ceramides, sphingosine, and its phosphorylated form sphingosine-1-phosphate are bioactive lipids that mediate cellular signaling pathways regulating several biological processes including cell proliferation, apoptosis and differentiation (PubMed : 10610716). Has a higher catalytic efficiency towards C12-ceramides versus other ceramides (PubMed : 15655246, PubMed : 7744740). Also catalyzes the reverse reaction allowing the synthesis of ceramides from fatty acids and sphingosine (PubMed : 12764132, PubMed : 12815059). For the reverse synthetic reaction, the natural sphingosine D-erythro isomer is more efficiently utilized as a substrate compared to D-erythro-dihydrosphingosine and D-erythro-phytosphingosine, while the fatty acids with chain lengths of 12 or 14 carbons are the most efficiently used (PubMed : 12764132). Has also an N-acylethanolamine hydrolase activity (PubMed : 15655246). By regulating the levels of ceramides, sphingosine and sphingosine-1-phosphate in the epidermis, mediates the calcium-induced differentiation of epidermal keratinocytes (PubMed : 17713573). Also indirectly regulates tumor necrosis factor/TNF-induced apoptosis (By similarity). By regulating the intracellular balance between ceramides and sphingosine, in adrenocortical cells, probably also acts as a regulator of steroidogenesis (PubMed : 22261821).. Isoform 2. May directly regulate steroidogenesis by binding the nuclear receptor NR5A1 and negatively regulating its transcriptional activity.
See full target information ASAH1
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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