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12666
Endosomal Marker Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Endosomal Marker Antibody Sampler Kit #12666

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Endosomal Marker Antibody Sampler Kit: Image 1

Flow cytometric analysis of HeLa cells using Caveolin-1 (D46G3) XP® Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Endosomal Marker Antibody Sampler Kit: Image 2

Confocal immunofluorescent analysis of SH-SY5Y cells using Clathrin Heavy Chain (D3C6) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Endosomal Marker Antibody Sampler Kit: Image 3

Confocal immunofluorescent analysis of HeLa cells using EEA1 (C45B10) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

Endosomal Marker Antibody Sampler Kit: Image 4

Confocal immunofluorescent analysis of HeLa cells using Rab5A (E6N8S) Mouse mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).

Endosomal Marker Antibody Sampler Kit: Image 5

Confocal immunofluorescent analysis of SK-MEL-28 cells using Rab7 (D95F2) XP® Rabbit mAb (green). Actin filaments were labeled using DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Endosomal Marker Antibody Sampler Kit: Image 6

Confocal immunofluorescent analysis of A549 cells using Rab11 (D4F5) XP® Rabbit mAb (green). Actin filaments were labeled wtih DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Endosomal Marker Antibody Sampler Kit: Image 7

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Endosomal Marker Antibody Sampler Kit: Image 8

Confocal immunofluorescent analysis of C2C12 cells using Caveolin-1 (D46G3) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).

Endosomal Marker Antibody Sampler Kit: Image 9

Western blot analysis of extracts from various cell lines using Clathrin Heavy Chain (D3C6) XP® Rabbit mAb.

Endosomal Marker Antibody Sampler Kit: Image 10

Western blot analysis of extracts from various cell lines using EEA1 (C45B10) Rabbit mAb.

Endosomal Marker Antibody Sampler Kit: Image 11

Immunohistochemical analysis of paraffin-embedded human ductal carcinoma of the breast using Rab5A (E6N8S) Mouse mAb.

Endosomal Marker Antibody Sampler Kit: Image 12

Western blot analysis of extracts from various cell lines using Rab7 (D95F2) XP® Rabbit mAb.

Endosomal Marker Antibody Sampler Kit: Image 13

Western blot analysis of extracts from various cell lines using Rab11 (D4F5) XP® Rabbit mAb.

Endosomal Marker Antibody Sampler Kit: Image 14

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Caveolin-1 (D46G3) XP® Rabbit mAb.

Endosomal Marker Antibody Sampler Kit: Image 15

Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using Rab5A (E6N8S) Mouse mAb.

Endosomal Marker Antibody Sampler Kit: Image 16

Immunohistochemical analysis of paraffin-embedded human lymphoma using Caveolin-1 (D46G3) XP® Rabbit mAb.

Endosomal Marker Antibody Sampler Kit: Image 17

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using Rab5A (E6N8S) Mouse mAb (left) compared to concentration matched Mouse (G3A1) IgG1 Isotype Control #5415 (right).

Endosomal Marker Antibody Sampler Kit: Image 18

Immunohistochemical analysis of paraffin-embedded mouse lung using Caveolin-1 (D46G3) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

Endosomal Marker Antibody Sampler Kit: Image 19

Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using Rab5A (E6N8S) Mouse mAb.

Endosomal Marker Antibody Sampler Kit: Image 20

Immunoprecipitation of caveolin-1 from HeLa cells using Caveolin-1 (D46G3) XP® Rabbit mAb followed by western blot using the same antibody. Lane 1 is 5% input.

Endosomal Marker Antibody Sampler Kit: Image 21

Immunoprecipitation of Rab5A protein from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Mouse (G3A1) mAb IgG1 Isotype Control #5415, lane 3 is Rab5A (E6N8S) Mouse mAb without HeLa cell extracts, and lane 4 is Rab5A (E6N8S) Mouse mAb. Western blot analysis was performed using Rab5A (E6N8S) Mouse mAb.

Endosomal Marker Antibody Sampler Kit: Image 22

Western blot analysis of extracts from various cell types using Caveolin-1 (D46G3) XP® Rabbit mAb.

Endosomal Marker Antibody Sampler Kit: Image 23

Immunoprecipitation of Rab5A protein from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is Rab5A (E6N8S) Mouse mAb. Western blot analysis was performed using Rab5 (C8B1) Rabbit mAb #3547.

Endosomal Marker Antibody Sampler Kit: Image 24

Western blot analysis of extracts from control HeLa cells (lane 1) or caveolin-1 knockout HeLa cells (lane 2) using Caveolin-1 (D46G3) XP® Rabbit mAb #3267 (upper), or β-actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the caveolin-1 knockout HeLa cells confirms the specificity of the antibody to caveolin-1.

Endosomal Marker Antibody Sampler Kit: Image 25

Western blot analysis of extracts from various cell lines using Rab5A (E6N8S) Mouse mAb.

To Purchase # 12666T
Product # Size Price
12666T
1 Kit  (6 x 20 µl) $ 445

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Caveolin-1 (D46G3) XP® Rabbit mAb 3267 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R Hm Mk B Dg 21, 24 Rabbit IgG
Clathrin Heavy Chain (D3C6) XP® Rabbit mAb 4796 20 µl
  • WB
  • IP
  • IF
H M R Mk 190 Rabbit IgG
EEA1 (C45B10) Rabbit mAb 3288 20 µl
  • WB
  • IP
  • IF
H M R 170 Rabbit IgG
Rab5A (E6N8S) Mouse mAb 46449 20 µl
  • WB
  • IP
  • IHC
  • IF
H M R Mk 25 Mouse IgG1
Rab7 (D95F2) XP® Rabbit mAb 9367 20 µl
  • WB
  • IP
  • IF
H M R Mk 23 Rabbit IgG
Rab11 (D4F5) XP® Rabbit mAb 5589 20 µl
  • WB
  • IP
  • IF
H M R Mk 25 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
  • WB
Horse 

Product Description

The Endosomal Marker Antibody Sampler Kit provides an economical means of distinguishing endosomes in the early, late, and recycling phases. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody will detect endogenous total levels of their target protein. The antibodies do not cross-react with other isoforms, with the exception of the Rab11 (D4F5) XP® Rabbit mAb which will detect both Rab11a and Rab11b.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu20 of human caveolin-1, Pro1663 of human clathrin heavy chain protein, Ser70 of human EEA1 protein, Gly190 of human Rab5A protein, Glu188 of human Rab7 protein, and near the amino terminus of human Rab11 protein.

Background

Endosomes are formed by the invagination of the plasma membrane to form vesicles in an effort to recycle components of the cell (1). Endosomes can be coated in clathrin when vesicles form at clathrin-coated pits (2). Caveolins are 21-24 kDa integral proteins that interact with cholesterol and are the main structural components of the cholesterol/sphingolipid-enriched plasma membrane caveolae (3). Each stage of endosome maturation is marked by a unique set of proteins. EEA1 is an early endosome marker that is essential for membrane fusion and trafficking (4). Members of the ras superfamily of small Rab GTPases, specifically Rab5, Rab7, and Rab11 are markers of the early, late and recycling endosomes (5).

  1. Huotari, J. and Helenius, A. (2011) EMBO J 30, 3481-500.
  2. Rodriguez-Boulan, E. et al. (2005) Nat Rev Mol Cell Biol 6, 233-47.
  3. Okamoto, T. et al. (1998) J Biol Chem 273, 5419-22.
  4. Christoforidis, S. et al. (1999) Nature 397, 621-625.
  5. Zerial, M. and McBride, H. (2001) Nat Rev Mol Cell Biol 2, 107-17.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
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.