Revision 1

#4753Store at -20C

1 Kit

(9 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
β-Tubulin (9F3) Rabbit mAb 2128 20 µl 55 kDa Rabbit IgG
COX IV (3E11) Rabbit mAb 4850 20 µl 17 kDa Rabbit IgG
NUP98 (C39A3) Rabbit mAb 2598 20 µl 98 kDa Rabbit IgG
Fibrillarin (C13C3) Rabbit mAb 2639 20 µl 37 kDa Rabbit IgG
LC3B (D11) XP® Rabbit mAb 3868 20 µl 14, 16 kDa Rabbit IgG
Rab5A (E6N8S) Mouse mAb 46449 20 µl 25 kDa Mouse IgG1
Calnexin (C5C9) Rabbit mAb 2679 20 µl 90 kDa Rabbit IgG
Histone H3 (D1H2) XP® Rabbit mAb 4499 20 µl 17 kDa Rabbit IgG
Histone H2A (D6O3A) Rabbit mAb 12349 20 µl 14 kDa Rabbit IgG

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

Description

The Cellular Localization IF Antibody Sampler Kit provides an economical means for identification of cellular organelles by fluorescence immnuocytochemistry (IF-IC). This kit includes enough primary antibody to perform at least twenty IF-IC tests or two Western blots with each 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

Knowledge of the subcellular location of a protein may reveal the potential role it plays in a variety of cellular processes. One can confirm the subcellular location of a marker that colocalizes with one of the organelle-specific antibodies in this kit. While these antibodies serve as powerful tools for immunofluorescence, they may also be used as western blot controls for fractionated cell lysates.

Background References

    Trademarks and Patents

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

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    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 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 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.

    Revision 1
    #4753

    Cellular Localization IF Antibody Sampler Kit

    Cellular Localization IF Antibody Sampler Kit: Image 1 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse liver using Fibrillarin (C13C3) Rabbit mAb (left, green), DyLight 554 Phalloidin #13054 (right, red), and DAPI #8961 (right, blue).
    Cellular Localization IF Antibody Sampler Kit: Image 2 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse kidney using Fibrillarin (C13C3) Rabbit mAb (left, green), DyLight 554 Phalloidin #13054 (right, red), and DAPI #8961 (right, blue).
    Cellular Localization IF Antibody Sampler Kit: Image 3 Expand Image
    Simple Western analysis of lysates (0.1 mg/mL) from HepG2 cells using Calnexin (C5C9) Rabbit mAb #2679. The virtual lane view (left) shows the target bands (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess ​​​​​​​Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    Cellular Localization IF Antibody Sampler Kit: Image 4 Expand Image

    Simple Western™ analysis of lysates (1mg/ml) from HeLa cells treated with Chloroquine (50uM, O/N) using LC3B (D11) XP® Rabbit mAb #3868. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 2-40kDa.

    Cellular Localization IF Antibody Sampler Kit: Image 5 Expand Image
    Simple Western analysis of lysates (1.0 mg/mL) from COS-7 cells using COX IV (3E11) Rabbit mAb #4850. The virtual lane view (left) shows the target bands (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess ​​​​​​​Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    Cellular Localization IF Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from various cell lines using Histone H2A (D6O3A) Rabbit mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from COS-7, NIH/3T3 and PC12 cells, using β-Tubulin (9F3) Rabbit mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of cell extracts from various cell types using NUP98 (C39A3) Rabbit mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from various cell types using Fibrillarin (C13C3) Antibody.
    Cellular Localization IF Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from PANC1, HepG2 and A204 cells using Calnexin (C5C9) Rabbit mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from HCT 116 and HCT 116 LC3B knockout cells, untreated (-) or treated with Chloroquine #14774 (50 μM, 18 hr) using LC3B (D11) XP® Rabbit mAb #3868 (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The absence of signal in the HCT 116 knockout cells confirms the specificity of the antibody for LC3B.
    Cellular Localization IF Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from various cell lines using Histone H3 (D1H2) XP® Rabbit mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 13 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Histone H3 (D1H2) XP® Rabbit mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 14 Expand Image
    Western blot analysis of extracts from various cell lines using Rab5A (E6N8S) Mouse mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 15 Expand Image
    Western blot analysis of extracts from HeLa, Jurkat and COS cell lines, using COX IV (3E11) Rabbit mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 16 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using Histone H2A (D6O3A) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
    Cellular Localization IF Antibody Sampler Kit: Image 17 Expand Image
    Immunohistochemical analysis of paraffin-embedded human glioblastoma using β-Tubulin (9F3) Rabbit mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 18 Expand Image
    Confocal immunofluorescent analysis of Caki-1 cells using NUP98 (C39A3) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
    Cellular Localization IF Antibody Sampler Kit: Image 19 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using Fibrillarin (C13C3) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
    Cellular Localization IF Antibody Sampler Kit: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Calnexin (C5C9) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
    Cellular Localization IF Antibody Sampler Kit: Image 21 Expand Image
    Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® LC3B siRNA I #6212 (+) or SignalSilence® LC3B siRNA II #6213 (+), using LC3B (D11) XP® Rabbit mAb #3868 and α-Tubulin (11H10) Rabbit mAb #2125. The LC3B (D11) XP® Rabbit mAb confirms silencing of LC3B expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of LC3B siRNA.
    Cellular Localization IF Antibody Sampler Kit: Image 22 Expand Image
    Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic mammary tumor using Histone H3 (D1H2) XP® Rabbit mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 23 Expand Image
    Western blot analysis of HAP1 extracts from WT (left) or RAB5A KO (right) using Rab5A (E6N8S) Mouse mAb. Membranes stained with Ponceau S for total protein normalization (lower). These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies, as a companion to validation data generated by CST scientists.
    Cellular Localization IF Antibody Sampler Kit: Image 24 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing staining of the mitochondria, using COX IV (3E11) Rabbit mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 25 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Histone H2A (D6O3A) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human MYT-1 Exon 1 Primers #4493, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
    Cellular Localization IF Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma using β-Tubulin (9F3) Rabbit mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 27 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using Calnexin (C5C9) Rabbit mAb (green). Actin filaments have been labeled using DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Cellular Localization IF Antibody Sampler Kit: Image 28 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using Histone H3 (D1H2) XP® Rabbit mAb (green) and β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red).
    Cellular Localization IF Antibody Sampler Kit: Image 29 Expand Image
    Immunohistochemical analysis of paraffin-embedded LL/2 syngeneic tumor using Histone H3 (D1H2) XP® Rabbit mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 30 Expand Image
    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.
    Cellular Localization IF Antibody Sampler Kit: Image 31 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using COX IV (3E11) Rabbit mAb in the presence of control peptide (left) or Cox IV Blocking Peptide #1034 (right).
    Cellular Localization IF Antibody Sampler Kit: Image 32 Expand Image
    Immunohistochemical analysis of paraffin-embedded human melanoma using β-Tubulin (9F3) Rabbit mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 33 Expand Image
    Flow cytometric analysis of HeLa cells using Histone H3 (D1H2) XP® Rabbit mAb (solid line) or a 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.
    Cellular Localization IF Antibody Sampler Kit: Image 34 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain using Histone H3 (D1H2) XP® Rabbit mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 35 Expand Image
    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.
    Cellular Localization IF Antibody Sampler Kit: Image 36 Expand Image
    Immunohistochemical analysis of paraffin-embedded H1650 xenograft, using COX IV Rabbit mAb. Note specific staining of human cancer cells.
    Cellular Localization IF Antibody Sampler Kit: Image 37 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using β-Tubulin (9F3) Rabbit mAb preincubated with control peptide (left) or β-Tubulin Blocking Peptide #1032 (right).
    Cellular Localization IF Antibody Sampler Kit: Image 38 Expand Image
    Immunohistochemical analysis of paraffin-embedded rhesus monkey liver using Histone H3 (D1H2) XP® Rabbit mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 39 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ductal carcinoma of the breast using Rab5A (E6N8S) Mouse mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 40 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using β-Tubulin (9F3) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Cellular Localization IF Antibody Sampler Kit: Image 41 Expand Image
    Confocal immunofluorescent analysis of HCT 116 cells either untreated (left) or treated with Chloroquine #14774 (50 µM, overnight) (center) or LC3B HCT 116 knockout cells treated with Chloroquine #14774 (50 µM, overnight) (right) using LC3B (D11) XP® Rabbit mAb (green). Actin filaments were labeled with β-Actin (8H10D10) Mouse mAb (red) and nuclei were labeled with DAPI #4083 (blue).
    Cellular Localization IF Antibody Sampler Kit: Image 42 Expand Image
    Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using Rab5A (E6N8S) Mouse mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 43 Expand Image
    Confocal immunofluorescent analysis of HeLa cells labeled with COX IV (3E11) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Cellular Localization IF Antibody Sampler Kit: Image 44 Expand Image
    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).
    Cellular Localization IF Antibody Sampler Kit: Image 45 Expand Image

    Flow cytometric analysis of K-562 cells using β-Tubulin (9F3) 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.

    Cellular Localization IF Antibody Sampler Kit: Image 46 Expand Image
    Flow cytometric analysis of HCT-116 cells, wild-type (green, high expression) or LC3B knockdown (blue, negative expression), using LC3B (D11) XP® Rabbit mAb #3868 (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    Cellular Localization IF Antibody Sampler Kit: Image 47 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using Rab5A (E6N8S) Mouse mAb.
    Cellular Localization IF Antibody Sampler Kit: Image 48 Expand Image
    Flow cytometric analysis of HeLa cells using COX IV (3E11) 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.
    Cellular Localization IF Antibody Sampler Kit: Image 49 Expand Image
    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).