Revision 7

#67955Store 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-P, IF-IC, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

55

Source/Isotype:

Rabbit IgG

UniProt ID:

#P20248

Entrez-Gene Id:

890

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:800 - 1:3200
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 #29113.

Specificity / Sensitivity

Cyclin A2 (E6D1J) XP® Rabbit mAb recognizes endogenous levels of total cyclin A2 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human cyclin A2 protein. The epitope corresponds to a region surrounding Glu121 of human cyclin A2.

Background

While overcoming the G1/S checkpoint to commence DNA replication requires cyclin E, and traversing the G2/M checkpoint to initiate mitosis requires cyclin B to be present, cyclin A seems to be required for both S-phase and M-phase (1). A number of studies have described the ability of overexpressed cyclin A to accelerate the G1 to S transition, causing DNA replication, and cyclin A antisense DNA can prevent DNA replication (2-4). Cyclin A availability is apparently the rate-limiting step for entry into mitosis, and cyclin A is required for the completion of prophase (5). At late prophase, cyclin A may no longer be necessary as cdc2/cyclinB1 becomes active (5).

  1. Pagano, M. et al. (1992) EMBO. J. 11, 961-71.
  2. Resnitzky, D. et al. (1995) Mol. Cell. Biol. 15, 4347-52.
  3. d'Urso, G. et al. (1990) Science 250, 786-91.
  4. Zindy, F. et al. (1992) Biochem. Biophys. Res. Commun. 182, 1144-54.
  5. Furuno, N. et al. (1999) J. Cell. Biol. 147, 295-306.

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-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.

使用に関する制限

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Revision 7
#67955

Cyclin A2 (E6D1J) XP® Rabbit mAb

Western Blotting Image 1: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various human cell lines, untreated (-) or treated with Aphidicolin #32774 (10 μg/mL, 24 hr; +) or Doxorubicin #5927 (0.5 μM, 24 hr; +), using Cyclin A2 (E6D1J) XP® Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Cyclin A2 protein is induced with aphidicolin and reduced with doxorubicin as expected. Low expression of cyclin A2 protein in Caki-1 cells is consistent with the predicted expression pattern.
Western Blotting Image 2: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from HCT 116 cells, transfected with control siRNA (-) or cyclin A2 siRNA (+), using Cyclin A2 (E6D1J) XP® Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Immunoprecipitation Image 1: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
Immunoprecipitation of cyclin A2 protein from HCT 116 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Cyclin A2 (E6D1J) XP® Rabbit mAb. Western blot analysis was performed using Cyclin A2 (E6D1J) XP® Rabbit mAb. Mouse Anti-Rabbit IgG (Light-Chain Specific) (D4W3E) mAb (HRP Conjugate) #93702 was used as the secondary antibody.
Immunohistochemistry Image 1: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using Cyclin A2 (E6D1J) XP® Rabbit mAb.
Immunohistochemistry Image 2: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Cyclin A2 (E6D1J) XP® Rabbit mAb.
Immunohistochemistry Image 3: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using Cyclin A2 (E6D1J) XP® Rabbit mAb.
Immunohistochemistry Image 4: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin lymphoma using Cyclin A2 (E6D1J) XP® Rabbit mAb.
Immunohistochemistry Image 5: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human large cell neuroendocrine carcinoma using Cyclin A2 (E6D1J) XP® Rabbit mAb.
Immunohistochemistry Image 6: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human small cell carcinoma of the salivary gland using Cyclin A2 (E6D1J) XP® Rabbit mAb.
Immunohistochemistry Image 7: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human thymus using Cyclin A2 (E6D1J) XP® Rabbit mAb.
Immunohistochemistry Image 8: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using Cyclin A2 (E6D1J) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right). 
Immunohistochemistry Image 9: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded HT-29 cell pellets, untreated (left) or treated with Aphidicolin #32774 (10 µg/ml, 24 hr; right), using Cyclin A2 (E6D1J) XP® Rabbit mAb.
Immunofluorescence Image 1: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of HCT 116 cells, either mock transfected (left, moderate-expressing), transfected with siRNA directed against human cyclin A2 (center, low-expressing), or treated with Aphidicolin #32774 (10 µg/mL, 24 hr; right, high-expressing), using Cyclin A2 (E6D1J) XP® Rabbit mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
Flow Cytometry Image 1: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
Flow cytometric analysis of Jurkat cells using DRAQ5® #4084 and Cyclin A2 (E6D1J) XP® Rabbit mAb (right) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 488® Conjugate) #4412 was used as a secondary antibody.