Revision 1

#9385Store at -20C

1 个试剂盒

(5 x 20 microliters)

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.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Rab4 Antibody 2167 20 µl 25 kDa Rabbit 
Rab5A (E6N8S) Mouse mAb 46449 20 µl 25 kDa Mouse IgG1
Rab7 (D95F2) XP® Rabbit mAb 9367 20 µl 23 kDa Rabbit IgG
Rab9A (D52G8) XP® Rabbit mAb 5118 20 µl 23 kDa Rabbit IgG
Rab11 (D4F5) XP® Rabbit mAb 5589 20 µl 25 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl Horse 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Rab Family Antibody Sampler Kit provides an economical means to evaluate the presence and status of Rab proteins in cells. This kit provides enough primary and secondary antibodies to perform two Western blot experiments per primary antibody.

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.

Background

Rab family proteins are GTPases and members of the Ras superfamily of monomeric G proteins. These membrane-associated proteins are involved in many aspects of vesicle-mediated transport, taking part in the initial vesicle formation, transport of vesicles along the cytoskeleton, and eventual fusion of vesicle and target membranes. Rab4 is localized at early endosomes/recycling endosomes and functions as a key regulator for sorting/recycling of membrane and proteins (1,2). Both Rab4A and Rab4B isoforms are localized to similar cellular compartments and are believed to have similar functions (4). Rab4 interacts with several Rab4 effectors in a complex on a special endosome site to promote membrane/protein recycling (1,3). Rab5 is localized to the plasma membrane and early endosome and functions as a key regulator of vesicle trafficking during early endocytosis (1). The conformational change between Rab5-GTP and Rab5-GDP is essential for its biological function as a rate-limiting regulator at multiple steps during endocytosis (1,5). Similar to Rab4, Rab5 also interacts with specific Rab5 effectors on a specialized endosomal Rab domain to promote recycling between endosome and the plasma membrane (1,5,6). Both Rab7 and Rab9 are located in late endosomes but exert different functions. Rab7 associates with the RIPL effector protein to control membrane trafficking from early to late endosome and to lysosomes (7,8). Rab7 also helps to regulate growth receptor endocytic trafficking and degradation, and maturation of phagosome and autophagic vacuoles (8-11). Rab9 interacts with its effector proteins p40 and TIP47 (12,13) to promote the MPR (mannose 6-phosphate receptor)-associated lysosomal enzyme transport between late endosomes and the trans Golgi network (14,15). Rab11 (isoforms Rab11a and Rab11b) functions as a key regulator in the recycling of perinuclear, plasma membrane and Golgi compartment endosomes (16,17). Despite some overlap, distinct differences exist between Rab11a and Rab11b in both their cellular distribution and functional roles. Rab11a is ubiquitously expressed while Rab11b is found mainly in the heart and brain (18,19). Like other Rab proteins, Rab11 functions when associated with Rab11 family interacting proteins (FIPs). The three distinct classes of Rab11 FIPs all share a conserved carboxy-terminal Rab-binding domain that allows Rab-FIP protein interaction. When bound together, these proteins are thought to regulate membrane-associated protein sorting (20,21).

  1. Zerial, M. and McBride, H. (2001) Nat Rev Mol Cell Biol 2, 107-17.
  2. van der Sluijs, P. et al. (1992) Cell 70, 729-40.
  3. Deneka, M. et al. (2003) EMBO J 22, 2645-57.
  4. Krawczyk, M. et al. (2007) Nucleic Acids Res 35, 595-605.
  5. van der Bliek, A.M. (2005) Nat Cell Biol 7, 548-50.
  6. Haas, A.K. et al. (2005) Nat Cell Biol 7, 887-93.
  7. Feng, Y. et al. (1995) J Cell Biol 131, 1435-52.
  8. Méresse, S. et al. (1995) J Cell Sci 108 ( Pt 11), 3349-58.
  9. Ceresa, B.P. and Bahr, S.J. (2006) J Biol Chem 281, 1099-106.
  10. Jäger, S. et al. (2004) J Cell Sci 117, 4837-48.
  11. Méresse, S. et al. (1999) EMBO J 18, 4394-403.
  12. Díaz, E. et al. (1997) J Cell Biol 138, 283-90.
  13. Barbero, P. et al. (2002) J Cell Biol 156, 511-8.
  14. Lombardi, D. et al. (1993) EMBO J 12, 677-82.
  15. Riederer, M.A. et al. (1994) J Cell Biol 125, 573-82.
  16. Ullrich, O. et al. (1996) J Cell Biol 135, 913-24.
  17. Chen, W. et al. (1998) Mol Biol Cell 9, 3241-57.
  18. Lapierre, L.A. et al. (2003) Exp Cell Res 290, 322-31.
  19. Khvotchev, M.V. et al. (2003) J Neurosci 23, 10531-9.
  20. Junutula, J.R. et al. (2004) J Biol Chem 279, 33430-7.
  21. Hales, C.M. et al. (2001) J Biol Chem 276, 39067-75.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    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 1
    #9385

    Rab Family Antibody Sampler Kit

    Rab Family Antibody Sampler Kit: Image 1 Expand Image
    使用 Rab4 Antibody 对不同细胞类型的提取物进行蛋白质印迹分析。
    Rab Family Antibody Sampler Kit: Image 2 Expand Image
    使用 Rab5A (E6N8S) 小鼠单克隆抗体对不同细胞系的提取物进行蛋白印迹分析。
    Rab Family Antibody Sampler Kit: Image 3 Expand Image
    使用 Rab9A (D52G8) XP® Rabbit mAb 对不同细胞系的提取物进行蛋白质印迹分析。
    Rab Family Antibody Sampler Kit: Image 4 Expand Image
    使用 Rab11 (D4F5) XP® Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
    Rab Family Antibody Sampler Kit: Image 5 Expand Image
    一抗与靶标蛋白结合之后,与偶联 HRP 的二抗形成复合体。添加 LumiGLO®,在酶催化分解期间发光。
    Rab Family Antibody Sampler Kit: Image 6 Expand Image
    一抗与靶标蛋白结合之后,与偶联 HRP 的二抗形成复合体。添加 LumiGLO*,它可在酶催化的分解期间发光。
    Rab Family Antibody Sampler Kit: Image 7 Expand Image
    使用 Rab7 (D95F2) XP® Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
    Rab Family Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of HAP1 extracts from WT (left) or RAB5A KO (right) using Rab5A (E6N8S) Mouse mAb. 用 Ponceau S 染色的膜进行总蛋白标准化(下图)。这些数据由 YCharOS Inc. 提供,该公司是一家开放科学公司,作为 CST 科学家生成的验证数据的合作伙伴,其使命是表征市售抗体。
    Rab Family Antibody Sampler Kit: Image 9 Expand Image
    使用 Rab9A (D52G8) XP® Rabbit mAb(绿色)对 HeLa 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 DY-554 Phalloidin 进行标记(红色)。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
    Rab Family Antibody Sampler Kit: Image 10 Expand Image
    使用 Rab11 (D4F5) XP® Rabbit mAb(绿色) 对 A549 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)。
    Rab Family Antibody Sampler Kit: Image 11 Expand Image
    使用 Rab7 (D95F2) XP® Rabbit mAb(绿色) 对 SK-MEL-28 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 DY-554 Phalloidin 进行标记(红色)。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
    Rab Family Antibody Sampler Kit: Image 12 Expand Image
    对 HeLa 细胞提取物 Rab5A 进行免疫沉淀。泳道 1 为 10% input,泳道 2 为Mouse (G3A1) mAb IgG1 Isotype Control #5415,泳道 3 为不含 HeLa 细胞提取物的 Rab5A (E6N8S) 小鼠单克隆抗体,泳道 4 为 Rab5A (E6N8S) 小鼠单克隆抗体。使用 Rab5A (E6N8S) 小鼠单克隆抗体进行蛋白印迹分析。
    Rab Family Antibody Sampler Kit: Image 13 Expand Image
    对 HeLa 细胞提取物 Rab5A 进行免疫沉淀。泳道 1 为 10% input,泳道 2 为Mouse (G3A1) mAb IgG1 Isotype Control #5415,泳道 3 为 Rab5A (E6N8S) 小鼠单克隆抗体。使用 Rab5 (C8B1) 兔单克隆抗体 #3547 进行蛋白印迹分析。
    Rab Family Antibody Sampler Kit: Image 14 Expand Image
    使用 Rab5A (E6N8S) 小鼠单克隆抗体对石蜡包埋的人乳腺导管癌进行免疫组织化学分析。
    Rab Family Antibody Sampler Kit: Image 15 Expand Image
    使用 Rab5A (E6N8S) 小鼠单克隆抗体对石蜡包埋的人卵巢浆液性乳头状癌进行免疫组织化学分析。
    Rab Family Antibody Sampler Kit: Image 16 Expand Image
    以浓度匹配的 Mouse (G3A1) IgG1 Isotype Control #5415(右图)作为对照,使用 Rab5A (E6N8S) 小鼠单克隆抗体(左图)对石蜡包埋的人结肠腺癌细胞进行免疫组织化学分析。
    Rab Family Antibody Sampler Kit: Image 17 Expand Image
    使用 Rab5A (E6N8S) 小鼠单克隆抗体对石蜡包埋的人前列腺腺癌进行免疫组织化学分析。
    Rab Family Antibody Sampler Kit: Image 18 Expand Image
    使用 Rab5A (E6N8S) 小鼠单克隆抗体(绿色)对 HeLa 细胞进行共聚焦免疫荧光分析。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)标记。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。