Revision 7

#93597

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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, W-S, IHC-P

Reactivity:
M R

Sensitivity:
Endogenous

MW (kDa):
30

Source/Isotype:
Rabbit IgG

UniProt ID:
#P30285

Entrez-Gene Id:
12567

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunohistochemistry (Paraffin) 1:50 - 1:200

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.

Specificity/Sensitivity

CDK4 (F8L5F) Rabbit Monoclonal Antibody recognizes endogenous levels of total CDK4 protein. Based on the sequence of the protein antigen, this antibody is not expected to detect other CDK proteins. This antibody detects approximately 12 kDa and 15 kDa proteins of unknown identity in some cell lines. This antibody is not recommended for immunohistochemical analysis of human tissues.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to full-length mouse CDK4 protein.

Background

Cyclin-dependent kinase activity is regulated by T-loop phosphorylation (Thr172 in the case of CDK4), by the abundance of their cyclin partners, and by association with CDK inhibitors of the Cip/Kip or INK family of proteins (1). The inactive ternary complex of CDK4/cyclin D and p27 Kip1/Cip1 requires extracellular mitogenic stimuli for the release and degradation of p27, which affects progression through the restriction point and pRb-dependent entry into S-phase (2). The active complex of CDK4/cyclin D targets the retinoblastoma protein for phosphorylation, allowing the release of E2F transcription factors that activate G1/S-phase gene expression (3). In HeLa cells, upon UV irradiation, upregulation of p16 INK4A association with CDK4/cyclin D3 leads to a G2 delay, implicating CDK4/cyclin D3 activity in progression through the G2-phase of the cell cycle (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 W-S: Simple Western™ IHC-P: Immunohistochemistry (Paraffin)

Cross-Reactivity Key

M: Mouse R: Rat

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.

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 7

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using CDK4 (F8L5F) Rabbit Monoclonal Antibody #93597 (upper) or GAPDH (D16H11) Rabbit Monoclonal Antibody #5174 (lower). Low expression of CDK4 protein in CTLL-2 cells is consistent with the predicted expression pattern.
Western Blotting Image 1: CDK4 (F8L5F) Rabbit Monoclonal Antibody
Western blot analysis of extracts from NMuMG cells, transfected with control siRNA (-) or mouse CDK4 siRNA (mCDK4 siRNA; +), using CDK4 (F8L5F) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 2: CDK4 (F8L5F) Rabbit Monoclonal Antibody
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length mouse CDK4 (mCDK4-Myc/DDK; +) using CDK4 (F8L5F) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 3: CDK4 (F8L5F) 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 7

Cell Signaling Technology Logo
Simple WesternTM analysis of lysates (1.0 mg/mL) from C2C12 cells CDK4 (F8L5F) Rabbit Monoclonal Antibody #93597. 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 JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western Blotting Image 1: CDK4 (F8L5F) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded mouse pancreas using CDK4 (F8L5F) Rabbit mAb.
Immunohistochemistry Image 1: CDK4 (F8L5F) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded rat liver using CDK4 (F8L5F) Rabbit mAb.
Immunohistochemistry Image 2: CDK4 (F8L5F) 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 7

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded mouse thymus using CDK4 (F8L5F) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 3: CDK4 (F8L5F) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic tumor using CDK4 (F8L5F) Rabbit mAb.
Immunohistochemistry Image 4: CDK4 (F8L5F) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded rat spleen using CDK4 (F8L5F) Rabbit mAb.
Immunohistochemistry Image 5: CDK4 (F8L5F) 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 7

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded mouse prostate using CDK4 (F8L5F) Rabbit mAb.
Immunohistochemistry Image 6: CDK4 (F8L5F) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded mouse spleen using CDK4 (F8L5F) Rabbit mAb.
Immunohistochemistry Image 7: CDK4 (F8L5F) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded mouse stomach using CDK4 (F8L5F) Rabbit mAb.
Immunohistochemistry Image 8: CDK4 (F8L5F) 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 7

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded LL/2 (LLC-1) syngeneic tumor using CDK4 (F8L5F) Rabbit mAb.
Immunohistochemistry Image 9: CDK4 (F8L5F) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded Renca syngeneic tumor using CDK4 (F8L5F) Rabbit mAb.
Immunohistochemistry Image 10: CDK4 (F8L5F) 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.