Revision 4

#23200

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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, IP, IHC-P, IF-F, IF-IC, FC-FP

Reactivity:
M

Sensitivity:
Endogenous

MW (kDa):
16

Source/Isotype:
Rabbit IgG

UniProt ID:
#P51480

Entrez-Gene Id:
12578

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:50 - 1:250
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:50 - 1:200
Immunofluorescence (Frozen) 1:1600
Immunofluorescence (Immunocytochemistry) 1:1600
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400

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 #44872.

Specificity/Sensitivity

p16 INK4A (F2T7H) Rabbit Monoclonal Antibody recognizes endogenous levels of total p16 INK4A protein. This antibody does not cross-react with p15 INK4B protein. Non-specific cytoplasmic staining was observed in mouse small intestine epithelium by immunohistochemistry.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to full-length mouse p16 INK4A protein. This antibody recognizes residues surrounding Arg156 of mouse p16 INK4A protein.

Background

Members of the INK4 family of cyclin-dependent kinase inhibitors include p16 INK4A, p15 INK4B, p18 INK4C, and p19 INK4D. The INK4 family members inhibit cyclin-dependent kinases 4 and 6 (CDK4 and CDK6), causing cell cycle arrest in G1 phase. The INK4A-ARF-INK4B locus on chromosome 9p21, frequently lost in human cancer, encodes the INK4 family members p16 INK5A and p15 INK4B, as well as the unrelated protein, ARF (1).

p16 INK4A expression, typically repressed in the absence of stress, is thought to drive cells into senescence, and p16 INK4A expression is a commonly used marker of senescent cells (2,3). p16 INK4A protein expression is often altered in human cancer, and high expression is currently used as a predictive biomarker in cervical cancer (3-5).

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

W: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

M: Mouse

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.

使用に関する制限

使用に関する制限

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研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

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 4

Cell Signaling Technologyロゴ
Western blot analysis of extracts from various cell lines using p16 INK4A (F2T7H) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Negative expression of p16 INK4A protein in NIH/3T3 cells is consistent with the predicted expression pattern.
Western Blotting Image 1: p16 INK4A (F2T7H) Rabbit Monoclonal Antibody
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing full-length mouse p16 INK4A protein (mINK4A-Myc/DDK; +) or full-length mouse p15 INK4B protein (mINK4B-Myc/DDK; +), using p16 INK4A (F2T7H) Rabbit mAb (upper) or Myc-Tag (71D10) Rabbit mAb #2278 (lower).
Western Blotting Image 2: p16 INK4A (F2T7H) Rabbit Monoclonal Antibody
Simple WesternTM analysis of lysates (0.1 mg/mL) from MEF WT cells using p16 INK4A (F2T7H) Rabbit mAb #23200. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody.  The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (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: p16 INK4A (F2T7H) 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 4

Cell Signaling Technologyロゴ
Immunoprecipitation of p16 INK4A protein from Renca cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is p16 INK4A (F2T7H) Rabbit mAb. Western blot analysis was performed using p16 INK4A (E5F3Y) Rabbit mAb #29271. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Immunoprecipitation Image 1: p16 INK4A (F2T7H) Rabbit Monoclonal Antibody

Immunohistochemical analysis of paraffin-embedded GL261 syngeneic tumor using p16 INK4A (F2T7H) Rabbit mAb.

Immunohistochemistry Image 1: p16 INK4A (F2T7H) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded A20 syngeneic tumor using p16 INK4A (F2T7H) Rabbit mAb.
Immunohistochemistry Image 2: p16 INK4A (F2T7H) 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 4

Cell Signaling Technologyロゴ

Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic tumor using p16 INK4A (F2T7H) Rabbit mAb.

Immunohistochemistry Image 3: p16 INK4A (F2T7H) Rabbit Monoclonal Antibody

Immunohistochemical analysis of paraffin-embedded mouse spleen using p16 INK4A (F2T7H) Rabbit mAb.

Immunohistochemistry Image 4: p16 INK4A (F2T7H) Rabbit Monoclonal Antibody

Immunohistochemical analysis of paraffin-embedded mouse thymus using p16 INK4A (F2T7H) Rabbit mAb.

Immunohistochemistry Image 5: p16 INK4A (F2T7H) 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 4

Cell Signaling Technologyロゴ
Immunohistochemical analysis of paraffin-embedded Renca syngeneic tumor using p16 INK4A (F2T7H) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 6: p16 INK4A (F2T7H) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded Renca cell pellet (left, positive) or NIH/3T3 cell pellet (right, negative) using p16 INK4A (F2T7H) Rabbit mAb.
Immunohistochemistry Image 7: p16 INK4A (F2T7H) Rabbit Monoclonal Antibody
Confocal immunofluorescent analysis of fixed frozen GL261 syngeneic tumor using p16 INK4A (F2T7H) Rabbit mAb (left) or no primary antibody (right) followed by detection with Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 (green). Sections were mounted in ProLong Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 1: p16 INK4A (F2T7H) 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 4

Cell Signaling Technologyロゴ
Confocal immunofluorescent analysis of Renca cells (left, positive) and C2C12 cells (right, negative) using p16 INK4A (F2T7H) Rabbit mAb (green), DyLight 554 Phalloidin #13054 (red), and DAPI #4083 (blue).
Immunofluorescence Image 1: p16 INK4A (F2T7H) Rabbit Monoclonal Antibody
Flow cytometric analysis of NIH/3T3 cells (blue, negative) and MEF cells (green, positive) using p16 INK4A (F2T7H) Rabbit mAb (solid lines) or 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.
Flow Cytometry Image 1: p16 INK4A (F2T7H) 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.