Asparagine synthetase Recombinant Rabbit Monoclonal Antibody [SC67-07]
Catalog# ET1610-50
Asparagine synthetase Recombinant Rabbit Monoclonal Antibody [SC67-07]
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WB
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IF-Cell
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IHC-P
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Human
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Mouse
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Rat
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Green monkey
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unconjugated
Overview
Product Name
Asparagine synthetase Recombinant Rabbit Monoclonal Antibody [SC67-07]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human Asparagine synthetase aa 502-538 / 561.
Species Reactivity
Human, Mouse, Rat, Green monkey
Validated Applications
WB, IF-Cell, IHC-P
Molecular Weight
Predicted band size: 64 kDa
Positive Control
K562 cell lysate, human skeletal muscle tissue lysate, human lung tissue lysates, K-562, mouse stomach tissue.
Conjugation
unconjugated
Clone Number
SC67-07
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000
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IF-Cell
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1:50-1:200
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IHC-P
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1:50-1:200
Target
Function
Asparagine synthetase (or aspartate-ammonia ligase) is a chiefly cytoplasmic enzyme that generates asparagine from aspartate. This amidation reaction is similar to that promoted by glutamine synthetase. The enzyme is ubiquitous in its distribution in mammalian organs, but basal expression is relatively low in tissues other than the exocrine pancreas. Cancerous cells exhibit rapid growth and cell division and subsequently have an increased nutritional need. The particularly low-level expression of asparagine synthetase in primary acute lymphoblastic leukemia (ALL) and numerous ALL cell lines, as compared to that of normal cells, makes asparagine depletion an effective method of treatment due to the cells' unusual dependency on circulating serum asparagine as a necessary nutrition for growth. A correlation between L-asparaginase efficacy and asparagine synthetase protein levels in a number of human ovarian cell lines has been observed. As mentioned above, this result confirmed similar observations in human leukemia cell lines. Hence asparagine synthetase might be used as a biomarker in ovarian cancer screening and potential treatment.
Background References
1. Huang ML et al. Molecular and functional alterations in a mouse cardiac model of Friedreich ataxia: activation of the integrated stress response, eIF2a phosphorylation, and the induction of downstream targets. Am J Pathol 183:745-57 (2013).
2. Scotti C et al. Cell-cycle inhibition by Helicobacter pylori L-asparaginase. PLoS One 5:e13892 (2010).
Subcellular Location
Cytoplasm.
Synonyms
asnS antibody
ASNS_HUMAN antibody
ASNSD antibody
Asparagine synthetase [glutamine-hydrolyzing] antibody
Asparagine synthetase antibody
Cell cycle control protein TS11 antibody
Glutamine dependent asparagine synthetase 3 antibody
Glutamine dependent asparagine synthetase antibody
Glutamine hydrolyzing antibody
Glutamine-dependent asparagine synthetase antibody
ExpandImages
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Western blot analysis of Asparagine synthetase on different lysates with Rabbit anti-Asparagine synthetase antibody (ET1610-50) at 1/2,000 dilution.
Lane 1: K-562 cell lysate (20 µg/Lane)
Lane 2: COS-1 cell lysate (20 µg/Lane)
Lane 3: Mouse brain tissue lysate (40 µg/Lane)
Lane 4: Rat brain tissue lysate (40 µg/Lane)
Predicted band size: 64 kDa
Observed band size: 55 kDa
Exposure time: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-50) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of K-562 cells labeling Asparagine synthetase with Rabbit anti-Asparagine synthetase antibody (ET1610-50) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Asparagine synthetase antibody (ET1610-50) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded mouse stomach tissue using anti-Asparagine synthetase antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-50, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"