Revision 6
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-Bond, IHC-P, FC-FP

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

64

Source/Isotype:

Rabbit IgG

UniProt ID:

#P08243

Entrez-Gene Id:

440

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
IHC Leica Bond 1:250 - 1:1000
Immunohistochemistry (Paraffin) 1:250 - 1:1000
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #81128.

Specificity / Sensitivity

Asparagine Synthetase (E6C2C) XP® Rabbit mAb recognizes endogenous levels of total asparagine synthetase protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys467 of human asparagine synthetase protein.

Background

Asparagine synthetase (ASNS) catalyzes the synthesis of asparagine from aspartate and glutamine. Research studies have shown that intracellular asparagine can suppress apoptosis in a large number of human tumors. In addition, ASNS expression levels have been associated with the progression of gliomas and neuroblastomas in patients (1). Furthermore, acute lymphocytic leukemia cells frequently depend upon serum asparagine for their viability, as they lack ASNS (2). Deprivation of asparagine by L-asparaginase has therefore been developed as a therapeutic treatment for acute lymphocytic leukemia (2,3). In subsets of gastric and hepatic cancers, ASNS promoter hypermethylation correlates with low ASNS expression, sensitizing these cancers to the asparaginase treatment (4).

  1. Zhang, J. et al. (2014) Mol Cell 56, 205-18.
  2. Gwinn, D.M. et al. (2018) Cancer Cell 33, 91-107.e6.
  3. Loayza-Puch, F. et al. (2016) Nature 530, 490-4.
  4. Li, H. et al. (2019) Nat Med 25, 850-60.
  5. Nakamura, A. et al. (2018) Proc Natl Acad Sci U S A 115, E7776-E7785.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 6
#92479

Asparagine Synthetase (E6C2C) XP® Rabbit mAb

Western Blotting Image 1: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
使用 Asparagine Synthetase (E6C2C) XP® Rabbit mAb(上图)和 GAPDH (D16H11) XP® Rabbit mAb #5174(下)对 K-562 和 Loucy 提取物进行蛋白质印迹分析。Loucy 细胞中缺乏可检测的天冬酰胺合成酶与已发表的数据(5)一致,证实了抗体对天冬酰胺合成酶的特异性。
Western Blotting Image 2: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
使用 Asparagine Synthetase (E6C2C) XP® Rabbit mAb(上图)和 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对 HCT 116、TALL-1 和 JHH-5 的提取物进行蛋白质印迹分析。TALL-1 和 JHH-5 细胞无法检测到天冬酰胺合成酶,分别与已发表的数据(5)和可公开获得的 RNAseq 数据一致,证实了抗体对天冬酰胺合成酶的特异性。
Immunoprecipitation Image 1: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
对 OVCAR-4 细胞提取物天冬酰胺合成酶蛋白进行免疫沉淀分析。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 Asparagine Synthetase (E6C2C) XP® Rabbit mAb。使用 Asparagine Synthetase (E6C2C) XP® Rabbit mAb 进行蛋白印迹分析。小鼠Anti-兔 IgG (轻链特异) (D4W3E) 单克隆抗体 (HRP 偶联) #93702 用作二抗。
Immunohistochemistry Image 1: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human oropharyngeal squamous cell carcinoma using Asparagine Synthetase (E6C2C) XP® Rabbit mAb performed on the Leica BOND RX.

Immunohistochemistry Image 2: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using Asparagine Synthetase (E6C2C) XP® Rabbit mAb performed on the Leica BOND RX.

Immunohistochemistry Image 3: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using Asparagine Synthetase (E6C2C) XP® Rabbit mAb performed on the Leica BOND RX.
Immunohistochemistry Image 1: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
使用 Asparagine Synthetase (E6C2C) XP® Rabbit mAb 对石蜡包埋的人非小细胞肺癌进行免疫组织化学分析。
Immunohistochemistry Image 2: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin lymphoma using Asparagine Synthetase (E6C2C) XP® Rabbit mAb.
Immunohistochemistry Image 3: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
使用 Asparagine Synthetase (E6C2C) XP® Rabbit mAb 对石蜡包埋的人结肠腺癌进行免疫组织化学分析。
Immunohistochemistry Image 4: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
使用 Asparagine Synthetase (E6C2C) XP® Rabbit mAb 对石蜡包埋的 JHH-5 异种移植物进行免疫组织化学分析。注意:小鼠免疫细胞中信号的存在和 ASNS 阴性 JHH-5 细胞中信号的缺乏。
Immunohistochemistry Image 5: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
使用 Asparagine Synthetase (E6C2C) XP® Rabbit mAb 对石蜡包埋的小鼠胰腺进行免疫组织化学分析。
Immunohistochemistry Image 6: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
使用 Asparagine Synthetase (E6C2C) XP® Rabbit mAb 对石蜡包埋的小鼠脾进行免疫组织化学分析。
Immunohistochemistry Image 7: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
使用 Asparagine Synthetase (E6C2C) XP® Rabbit mAb(左图)或 Asparagine Synthetase Rabbit mAb(右图)对石蜡包埋的人非小细胞肺癌进行免疫组织化学分析。这两种抗体可检测人天冬酰胺合成酶上的独立、独特表位。使用两种抗体获得的相似染色模式有助于确认染色特异性。
Immunohistochemistry Image 8: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(右图)作为对照,使用 Asparagine Synthetase (E6C2C) XP® Rabbit mAb(左图)对石蜡包埋的人胰腺进行免疫组织化学分析。
Immunohistochemistry Image 9: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
使用 Asparagine Synthetase (E6C2C) XP® Rabbit mAb 对石蜡包埋的 HCT 细胞沉淀物(左图,阳性)或 HCT 116 细胞沉淀物(右图,阴性)进行免疫组织化学分析。
Flow Cytometry Image 1: Asparagine Synthetase (E6C2C) XP® Rabbit mAb Expand Image
使用 Asparagine Synthetase (E6C2C) XP® Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对 Loucy 细胞(蓝色)和 K-562 细胞(绿色)进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。