Revision 1
Cell Signaling Technology

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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
ESET (C1C12) Rabbit mAb 2196 40 µl 180 kDa Rabbit IgG
Fibrillarin (C13C3) Rabbit mAb 2639 40 µl 37 kDa Rabbit IgG
Histone H2A.Z Antibody 2718 40 µl 14 kDa Rabbit 
Histone H3 (D1H2) XP® Rabbit mAb 4499 40 µl 17 kDa Rabbit IgG
Lamin A/C (4C11) Mouse mAb 4777 40 µl 74 (Lamin A), 63 (Lamin C) kDa Mouse IgG2a
LSD1 (C69G12) Rabbit mAb 2184 40 µl 110 kDa Rabbit IgG
NUP98 (C39A3) Rabbit mAb 2598 40 µl 98 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 Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit provides an economical means to evaluate relevant nuclear proteins. This kit contains enough primary antibody to perform at least four western blots 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

The Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit contains a variety of antibodies directed against established nuclear proteins (1). Histone H3 and histone H2A.Z are histone family members and components of nucleosomes, the primary building block of chromatin made up of DNA wound around eight core histone proteins. The amino-terminal tails of core histones undergo various post-translational modifications and have a direct effect on the accessibility of chromatin to transcription factors and, therefore, gene expression (2). ESET histone methyltransferase (3) and LSD1 histone demethylase (4) are both regulators of histone methylation and are chromatin-associated. Both NUP98 (5) and lamins (6) are located within the nuclear envelope (also known as the nuclear membrane). NUP98 is a component of the nuclear pore complex. Lamin A and lamin C are fibrous proteins contributing to nuclear structural and transcriptional regulation. Finally, fibrillarin (7) is located in fibrillar regions and Cajal bodies of nucleoli, where it functions to regulate RNA transcription and pre-rRNA processing.

  1. Workman, J.L. and Kingston, R.E. (1998) Annu Rev Biochem 67, 545-79.
  2. Jin, J. et al. (2005) Trends Biochem Sci 30, 680-7.
  3. Yang, L. et al. (2002) Oncogene 21, 148-52.
  4. Shi, Y. et al. (2004) Cell 119, 941-53.
  5. Fontoura, B.M. et al. (1999) J Cell Biol 144, 1097-112.
  6. Gruenbaum, Y. et al. (2000) J Struct Biol 129, 313-23.
  7. Tollervey, D. et al. (1993) Cell 72, 443-57.

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.

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    Revision 1
    #12784

    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit

    Nucleus and Nuclear Envelope-Associated Marker Proteins 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).
    Nucleus and Nuclear Envelope-Associated Marker Proteins 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).
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 3 Expand Image
    Flow cytometric analysis of Daudi cells (blue) and MJ cells (green) using Lamin A/C (4C11) Mouse mAb (solid lines) or concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 4 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from HeLa cells using Lamin A/C (4C11) Mouse mAb #4777. 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.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of cell lysates from various cell types using LSD1 (C69G12) Rabbit mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of cell lysates from MCF7 and 293 cells using ESET (C1C12) Rabbit mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of cell extracts from various cell types using NUP98 (C39A3) Rabbit mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from various cell types using Fibrillarin (C13C3) Antibody.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of HeLa, NIH/3T3, H-4-II-E and COS cell lysates, in addition to 10 ng of recombinant H2A (rH2A) and H2A.Z (rH2A.Z) protein, using Histone H2A.Z Antibody #2718 (upper) and Histone H2A Antibody II #2578 (lower).
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from various cell lines using Histone H3 (D1H2) XP® Rabbit mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 11 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Histone H3 (D1H2) XP® Rabbit mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from control HeLa cells (lane 1) or HeLa cells with an apparent in-frame truncation mutation in the gene encoding LMNA (lane 2) using Lamin A/C (4C11) Mouse mAb #4777 (upper) or α-actinin (D6F6) XP® Rabbit mAb #6487 (lower). The change in LMNA molecular weight in the mutated HeLa cells is consistent with an in-frame deletion.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 13 Expand Image
    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.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 14 Expand Image
    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.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 15 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using LSD1 (C69G12) Rabbit mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 16 Expand Image
    Confocal immunofluorescent analysis of MCF-7 cells using ESET (C1C12) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 17 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).
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 18 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).
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 19 Expand Image
    Confocal immunofluorescent analysis of HeLa cells, using Histone H2A.Z Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic mammary tumor using Histone H3 (D1H2) XP® Rabbit mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 21 Expand Image
    Western blot analysis of extracts from various cell lines using Lamin A/C (4C11) Mouse mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 22 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma using LSD1 (C69G12) Rabbit mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 23 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).
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 24 Expand Image
    Immunohistochemical analysis of paraffin-embedded LL/2 syngeneic tumor using Histone H3 (D1H2) XP® Rabbit mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 25 Expand Image
    Western blot analysis of extracts from THP-1 cells, untreated or treated with cycloheximide (CHX, 10 μg/ml, overnight) followed by TNF-α #8902 (20 ng/ml, 4 hr), using Lamin A/C (4C11) Mouse mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded human stomach carcinoma using LSD1 (C69G12) Rabbit mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 27 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.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 28 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain using Histone H3 (D1H2) XP® Rabbit mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 29 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Lamin A/C (4C11) Mouse mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 30 Expand Image
    Immunohistochemical analysis of paraffin-embedded rhesus monkey liver using Histone H3 (D1H2) XP® Rabbit mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 31 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Lamin A/C (4C11) Mouse mAb.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 32 Expand Image
    Immunofluorescent analysis of normal rat brain using Lamin A/C (4C11) Mouse mAb (green) and MAP2 Antibody #4542 (red).
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 33 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using LSD1 (C69G12) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 34 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using Lamin A/C (4C11) Mouse mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 35 Expand Image
    Flow cytometric analysis of HeLa cells (green) using Lamin A/C (4C11) Mouse mAb (solid lines) or a concentration matched Mouse (G3A1) mAb IgG Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
    Nucleus and Nuclear Envelope-Associated Marker Proteins Antibody Sampler Kit: Image 36 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from U-937 cells and either LSD1 (C69G12) 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 SimpleChIP® Human CD11b Promoter Primers #14271, human SPI-1 promoter primers, 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.