Revision 3

#59286Store at -20C

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, IF-IC, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

15

Source/Isotype:

Rabbit IgG

UniProt ID:

#P22749

Entrez-Gene Id:

10578

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:800 - 1:3200
Flow Cytometry (Fixed/Permeabilized) 1:400 - 1:1600

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 #91378.

Specificity / Sensitivity

GNLY (E1V5M) Rabbit mAb recognizes endogenous levels of total GNLY protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human GNLY protein.

Background

Granulysin (GNLY) was originally identified as a late activation marker in T cells, and it is expressed by killer lymphocytes in most mammals, but not rodents. GNLY is largely confined to cytotoxic granules but can be secreted by killer cells, especially those expressing high levels of GNLY, such as decidual natural killer cells (1-3). GNLY is produced as a 15 kDa protein and is processed into a 9 kDa active pore-forming fragment by proteolytic removal of peptides from both the N- and C-termini. Moreover, the 15 kDa form was reported to function as an immune alarmin, causing the maturation and migration of antigen-presenting cells and other cells of the immune system (4-7). Unlike perforin, a cholesterol-dependent pore-forming protein that preferentially permeabilizes mammalian membranes, GNLY is inhibited by cholesterol and forms pores much more efficiently in microbial than mammalian membranes, and it plays an important role against bacteria, fungi, and parasite infections (8,9).

  1. Jongstra, J. et al. (1987) J Exp Med 165, 601-14.
  2. Pitabut, N. et al. (2011) Microbiol Immunol 55, 565-73.
  3. Crespo, Â.C. et al. (2020) Cell 182, 1125-1139.e18.
  4. Hanson, D.A. et al. (1999) Mol Immunol 36, 413-22.
  5. Tewary, P. et al. (2010) Blood 116, 3465-74.
  6. Clayberger, C. et al. (2012) J Immunol 188, 6119-26.
  7. Sparrow, E. and Bodman-Smith, M.D. (2020) Immunol Lett 217, 126-132.
  8. Kumar, J. et al. (2001) Expert Opin Investig Drugs 10, 321-9.
  9. Dotiwala, F. and Lieberman, J. (2019) Curr Opin Immunol 60, 19-29.

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 IF-IC: Immunofluorescence (Immunocytochemistry) 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 3
#59286

GNLY (E1V5M) Rabbit mAb

Western Blotting Image 1: GNLY (E1V5M) Rabbit mAb Expand Image
使用 GNLY (E1V5M) Rabbit mAb(上图)和 β-Actin (D6A8) Rabbit mAb #(下图)对各种细胞系、富集的人外周血单核细胞 CD56+ 自然杀伤 (NK) 细胞和活化的人 CD8+8457 T 细胞进行蛋白质印迹分析。
Immunoprecipitation Image 1: GNLY (E1V5M) Rabbit mAb Expand Image
对 NK-92 细胞提取物的 GNLY 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 GNLY (E1V5M) Rabbit mAb。使用 GNLY (E1V5M) Rabbit mAb 进行蛋白质印迹分析。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 用作二抗。
Immunofluorescence Image 1: GNLY (E1V5M) Rabbit mAb Expand Image
使用 GNLY (E1V5M) Rabbit mAb(绿色)、DyLight 554 Phalloidin #13054(红色)和 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)对 NK-92 细胞(左图,阳性)和 K-562 细胞(右图,阴性)进行共聚焦免疫荧光分析。
Flow Cytometry Image 1: GNLY (E1V5M) Rabbit mAb Expand Image
使用 GNLY (E1V5M) Rabbit mAb(右图)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(左图)和共染色的 NCAM1 (CD56) (MY31) Mouse mAb (APC Conjugate) #51997 对固定和透化的人外周血单核细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。分析对象为淋巴细胞门中的细胞。