Revision 5

#55506Store at -20C

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.
Applications:

WB, IP, IHC-Bond, IHC-P, IF-IC, FC-FP

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

36

Source/Isotype:

Rabbit IgG

UniProt ID:

#P24385

Entrez-Gene Id:

595

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
IHC Leica Bond 1:250 - 1:1000
Immunohistochemistry (Paraffin) 1:250 - 1:1000
Immunofluorescence (Immunocytochemistry) 1:400 - 1:1600
Flow Cytometry (Fixed/Permeabilized) 1:400 - 1:1600

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.

For a carrier free (BSA and azide free) version of this product see product #66467.

Specificity / Sensitivity

Cyclin D1 (E3P5S) XP® Rabbit mAb recognizes endogenous levels of total cyclin D1 protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala284 of human cyclin D1 protein.

Background

Activity of the cyclin-dependent kinases CDK4 and CDK6 is regulated by T-loop phosphorylation, by the abundance of their cyclin partners (the D-type cyclins), and by association with CDK inhibitors of the Cip/Kip or INK family of proteins (1). The inactive ternary complex of cyclin D/CDK4 and p27 Kip1 requires extracellular mitogenic stimuli for the release and degradation of p27 concomitant with a rise in cyclin D levels to affect progression through the restriction point and Rb-dependent entry into S-phase (2). The active complex of cyclin D/CDK4 targets the retinoblastoma protein for phosphorylation, allowing the release of E2F transcription factors that activate G1/S-phase gene expression (3). Levels of cyclin D protein drop upon withdrawal of growth factors through downregulation of protein expression and phosphorylation-dependent degradation (4).

  1. Hirai, H. et al. (1995) Mol Cell Biol 15, 2672-81.
  2. Sherr, C.J. (1996) Science 274, 1672-7.
  3. Lukas, J. et al. (1996) Mol Cell Biol 16, 6917-25.
  4. Diehl, J.A. et al. (1997) Genes Dev 11, 957-72.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
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 5
#55506

Cyclin D1 (E3P5S) XP® Rabbit mAb

Western Blotting Image 1: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using Cyclin D1 (E3P5S) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation Image 1: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
Immunoprecipitation of Cyclin D1 from SH-SY5Y cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Cyclin D1 (E3P5S) XP® Rabbit mAb. Western blot analysis was performed usingCyclin D1 (E3P5S) XP® Rabbit mAb as the primary antibody and Anti-rabbit IgG, HRP-linked Antibody #7074 as the secondary antibody.
Immunohistochemistry Image 1: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using Cyclin D1 (E3P5S) XP® Rabbit mAb performed on the Leica® BOND Rx. 
Immunohistochemistry Image 1: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using Cyclin D1 (E3P5S) XP® Rabbit mAb.
Immunohistochemistry Image 2: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using Cyclin D1 (E3P5S) XP® Rabbit mAb (left) or Cyclin D1 (92G2) Rabbit mAb #2978 (right). These two antibodies detect independent, unique epitopes on human cyclin D1. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Immunohistochemistry Image 3: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using Cyclin D1 (E3P5S) XP® Rabbit mAb.
Immunohistochemistry Image 4: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded 4T1 metastatic tumor in mouse lung using Cyclin D1 (E3P5S) XP® Rabbit mAb.
Immunohistochemistry Image 5: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ovarian carcinosarcoma using Cyclin D1 (E3P5S) XP® Rabbit mAb.
Immunohistochemistry Image 6: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse pancreas using Cyclin D1 (E3P5S) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 7: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded SH-SY5Y cell pellet (left, positive) or THP-1 cell pellet (right, negative) using Cyclin D1 (E3P5S) XP® Rabbit mAb.
Immunofluorescence Image 1: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of SH-SY5Y cells (left, positive) and 293T cells (right, negative) using Cyclin D1 (E3P5S) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red).
Flow Cytometry Image 1: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
Flow cytometric analysis of K-562 cells (blue) and SH-SY5Y cells (green) using Cyclin D1 (E3P5S) XP® Rabbit mAb (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.