Revision 3

#52508

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IP, IHC-P, IF-IC

Reactivity:
H Mk

Sensitivity:
Endogenous

MW (kDa):
25

Source/Isotype:
Rabbit IgG

UniProt ID:
#O75496

Entrez-Gene Id:
51053

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:200 - 1:800
Immunofluorescence (Immunocytochemistry) 1:800 - 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 #44203.

Specificity/Sensitivity

Geminin (E5Q9S) Rabbit Monoclonal Antibody recognizes endogenous levels of total geminin protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro70 of human geminin protein.

Background

The initiation of DNA replication in mammalian cells is a highly coordinated process that ensures duplication of the genome only once per cell division cycle. Origins of replication are dispersed throughout the genome and their activities are regulated via the sequential binding of pre-replication and replication factors. The origin recognition complex (ORC) is thought to be bound to chromatin throughout the cell cycle (1,2). The pre-replication complex (pre-RC) forms in late mitosis/early G1 phase beginning with the binding of CDT1 and cdc6 to the origin, which allows binding of the heterohexameric MCM2-7 complex. Once this complex is formed, the origin is “licensed” for initiation of DNA replication. In order to ensure that replication occurs only once per cell cycle, geminin binds to and inhibits CDT1 during the S, G2, and M phases. This prevents the recruitment of the MCM complex to the origins of replication, which blocks the premature reformation of the pre-RC. At the metaphase/anaphase transition, geminin is degraded by the anaphase-promoting complex (APC) allowing for the formation of a new pre-RC (3,4).

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: Human Mk: Monkey

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

SignalStain is a registered 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.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Western bot analysis of extracts from various cell lines using Geminin (E5Q9S) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 1: Geminin (E5Q9S) Rabbit Monoclonal Antibody
Immunoprecipitation of Geminin from 293T cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Geminin (E5Q9S) XP® Rabbit mAb. Western blot analysis was performed using

Geminin (E5Q9S) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.

Immunoprecipitation Image 1: Geminin (E5Q9S) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded 293T cell pellet (left, high-expressing) or SK-MEL-28 cell pellet (right, low-expressing) using Geminin (E5Q9S) XP® Rabbit mAb.
Immunohistochemistry Image 1: Geminin (E5Q9S) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using Geminin (E5Q9S) XP® Rabbit mAb.
Immunohistochemistry Image 2: Geminin (E5Q9S) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded human esophageal carcinoma using Geminin (E5Q9S) XP® Rabbit mAb.
Immunohistochemistry Image 3: Geminin (E5Q9S) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using Geminin (E5Q9S) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 4: Geminin (E5Q9S) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using Geminin (E5Q9S) XP® Rabbit mAb.
Immunohistochemistry Image 5: Geminin (E5Q9S) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin's lymphoma using Geminin (E5Q9S) XP® Rabbit mAb.
Immunohistochemistry Image 6: Geminin (E5Q9S) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using Geminin (E5Q9S) XP® Rabbit mAb.
Immunohistochemistry Image 7: Geminin (E5Q9S) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of Hep G2 cells (left) and SK-MEL-28 cells (right) using Geminin (E5Q9S) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 1: Geminin (E5Q9S) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.