Revision 6

#11948Store 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:

M

SENSITIVITY:

Endogenous

MW (kDa):

17,25,28

Source/Isotype:

Rabbit IgG

UniProt ID:

#P06804

Entrez-Gene Id:

21926

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:100 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400

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 #57485.

Specificity / Sensitivity

TNF-α (D2D4) XP® Rabbit mAb recognizes endogenous levels of total TNF-α protein.

Species Reactivity:

Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a recombinant mouse TNF-α protein.

Background

TNF-α, the prototypical member of the TNF protein superfamily, is a homotrimeric type-II membrane protein (1,2). Membrane-bound TNF-α is cleaved by the metalloprotease TACE/ADAM17 to generate a soluble homotrimer (2). Both membrane and soluble forms of TNF-α are biologically active. TNF-α is produced by a variety of immune cells including T cells, B cells, NK cells, and macrophages (1). Cellular response to TNF-α is mediated through interaction with receptors TNF-R1 and TNF-R2 and results in activation of pathways that favor both cell survival and apoptosis depending on the cell type and biological context. Activation of kinase pathways (including JNK, Erk1/2, p38 MAPK, and NF-κB) promotes the survival of cells, while TNF-α-mediated activation of caspase-8 leads to programmed cell death (1,2). TNF-α plays a key regulatory role in inflammation and host defense against bacterial infection, notably Mycobacterium tuberculosis (3).
Mouse and rat TNF-α are predicted to be glycosylated, while human TNF-α is not (4).

  1. Aggarwal, B.B. (2003) Nat Rev Immunol 3, 745-56.
  2. Hehlgans, T. and Pfeffer, K. (2005) Immunology 115, 1-20.
  3. Lin, P.L. et al. (2007) J Investig Dermatol Symp Proc 12, 22-5.
  4. Pennica, D. et al. (1985) Proc Natl Acad Sci U S A 82, 6060-4.

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 BSA, 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.
XP is a registered trademark of Cell Signaling Technology, Inc.
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
#11948

TNF-α (D2D4) XP® Rabbit mAb

Western Blotting Image 1: TNF-α (D2D4) XP® Rabbit mAb Expand Image
使用 TNF-α (D2D4) XP® Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb#8457(下图),对未经处理 (-) 或经过 LPS(100 ng/mL,6 小时)和 Brefeldin A #9972(300 ng/mL,持续刺激 3 小时)处理的 Raw 264.7 细胞提取物进行蛋白质印迹法分析。
Western Blotting Image 2: TNF-α (D2D4) XP® Rabbit mAb Expand Image
使用 TNF-α (D2D4) XP® Rabbit mAb 对 1 ng 重组小鼠  TNF-α #5178 进行蛋白质印迹法分析。
Immunoprecipitation Image 1: TNF-α (D2D4) XP® Rabbit mAb Expand Image
使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(泳道 2)或 TNF-α (D2D4) XP® Rabbit mAb(泳道 3),对经过 LPS(100 ng/mL,6 小时)和 Brefeldin A #9972(300 ng/mL,持续刺激 3 小时)处理的 Raw 264.7 细胞提取物进行免疫沉淀分析。泳道 1 是 10% 输入对照。使用 TNF-α (D2D4) XP® Rabbit mAb 进行蛋白质印迹法分析。使用 Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) #3678 与 Anti-mouse IgG, HRP-linked Antibody #7076ntibody #作为二抗。
Immunofluorescence Image 1: TNF-α (D2D4) XP® Rabbit mAb Expand Image
使用 TNF-α (D2D4) XP® Rabbit mAb(绿色)对经过 LPS(100 ng/ml,6 小时;左图)处理或未经处理(右图)的 Raw 264.7 细胞进行共聚焦免疫荧光分析。使用 DY-554 phalloidin(红色)标记肌动蛋白纤丝。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
Flow Cytometry Image 1: TNF-α (D2D4) XP® Rabbit mAb Expand Image
使用 TNF-α (D2D4) XP® Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对未经处理(蓝色)或经过 LPS(100 ng/ml,6 小时)和 Brefeldin A #9972(300 ng/ml,持续处理 3 小时)处理(绿色)的 Raw 264.7 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。