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.

    使用に関する制限

    法的な権限を与えられたCSTの担当者が署名した書面によって別途明示的に合意された場合を除き、 CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、 または、ここに定められた条件と異なるものは、法的な権限を与えられたCSTの担当者が別途書面にて受諾した場合を除き、拒絶され、 いかなる効力も効果も有しません。

    研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

    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).