Anti-Cyclin D1 抗体 [SP4] (ab16663)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [SP4] to Cyclin D1
- Suitable for: ICC/IF, Flow Cyt (Intra), WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Cyclin D1 antibody [SP4]
Cyclin D1 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [SP4] to Cyclin D1 -
由来種
Rabbit -
アプリケーション
適用あり: ICC/IF, Flow Cyt (Intra), WB, IHC-Pmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is considered to be commercially sensitive.
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エピトープ
C-terminus -
ポジティブ・コントロール
- WB: MCF7, Hap1, A431 and HeLa, Nuero-2a, NIH/3T3, C6, Wild-type A549 and SH-SY5Y cell lysates. IHC (FFPE): Human normal tonsil; breast carcinoma; mantle cell lymphoma; rat esophagus. ICC/IF: MCF7 cells, C6, Neuro-2a and HAP1 cells (HAP1-CCND1 knockout cells used as negative cell line). Flow Cyt (intra): MCF7, NIH/3T3 and C6 cells.
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特記事項
This product was switched from a hybridoma to recombinant production method on 22nd January 2019.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles. -
バッファー
pH: 7.20
Preservative: 0.1% Sodium azide
Constituents: 1% BSA, PBS -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
SP4 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab16663の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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ICC/IF | (5) |
1/50 - 1/250.
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Flow Cyt (Intra) |
1/30.
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WB | (12) |
1/25 - 1/200. Detects a band of approximately 36 kDa (predicted molecular weight: 33 kDa).
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IHC-P | (5) |
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Deparaffinization: Deparaffinize slides using xylene or xylene alternative and graded alcohols. Antigen Retrieval: Boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at room temperature for 20 min. Primary Antibody Incubation: Incubate for 30 minutes at room temperature. Slide Washing: Slides must be washed in between steps. Rinse slides with PBS/0.05% Tween. |
特記事項 |
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ICC/IF
1/50 - 1/250. |
Flow Cyt (Intra)
1/30. |
WB
1/25 - 1/200. Detects a band of approximately 36 kDa (predicted molecular weight: 33 kDa). |
IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Deparaffinization: Deparaffinize slides using xylene or xylene alternative and graded alcohols. Antigen Retrieval: Boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at room temperature for 20 min. Primary Antibody Incubation: Incubate for 30 minutes at room temperature. Slide Washing: Slides must be washed in between steps. Rinse slides with PBS/0.05% Tween. |
ターゲット情報
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機能
Essential for the control of the cell cycle at the G1/S (start) transition. -
関連疾患
Note=A chromosomal aberration involving CCND1 may be a cause of B-lymphocytic malignancy, particularly mantle-cell lymphoma (MCL). Translocation t(11;14)(q13;q32) with immunoglobulin gene regions. Activation of CCND1 may be oncogenic by directly altering progression through the cell cycle.
Note=A chromosomal aberration involving CCND1 may be a cause of parathyroid adenomas. Translocation t(11;11)(q13;p15) with the parathyroid hormone (PTH) enhancer.
Defects in CCND1 are a cause of multiple myeloma (MM) [MIM:254500]. MM is a malignant tumor of plasma cells usually arising in the bone marrow and characterized by diffuse involvement of the skeletal system, hyperglobulinemia, Bence-Jones proteinuria and anemia. Complications of multiple myeloma are bone pain, hypercalcemia, renal failure and spinal cord compression. The aberrant antibodies that are produced lead to impaired humoral immunity and patients have a high prevalence of infection. Amyloidosis may develop in some patients. Multiple myeloma is part of a spectrum of diseases ranging from monoclonal gammopathy of unknown significance (MGUS) to plasma cell leukemia. Note=A chromosomal aberration involving CCND1 is found in multiple myeloma. Translocation t(11;14)(q13;q32) with the IgH locus. -
配列類似性
Belongs to the cyclin family. Cyclin D subfamily. -
翻訳後修飾
Phosphorylation at Thr-286 by MAP kinases is required for ubiquitination and degradation following DNA damage. It probably plays an essential role for recognition by the FBXO31 component of SCF (SKP1-cullin-F-box) protein ligase complex.
Ubiquitinated, primarily as 'Lys-48'-linked polyubiquitination. Ubiquitinated by a SCF (SKP1-CUL1-F-box protein) ubiquitin-protein ligase complex containing FBXO4 and CRYAB (By similarity). Following DNA damage it is ubiquitinated by some SCF (SKP1-cullin-F-box) protein ligase complex containing FBXO31. Ubiquitination leads to its degradation and G1 arrest. Deubiquitinated by USP2; leading to stabilize it. -
細胞内局在
Nucleus. - Information by UniProt
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参照データベース
- Entrez Gene: 595 Human
- Entrez Gene: 12443 Mouse
- Entrez Gene: 58919 Rat
- Omim: 168461 Human
- SwissProt: P24385 Human
- SwissProt: P25322 Mouse
- SwissProt: P39948 Rat
- Unigene: 523852 Human
see all -
別名
- AI327039 antibody
- B cell CLL/lymphoma 1 antibody
- B cell leukemia 1 antibody
see all
画像
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All lanes : Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/25 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : ccnd1 knockout A549 cell lysate
Lane 3 : SH-SY5Y cell lysate
Lane 4 : K562 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 33 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Cyclin D1 antibody [SP4] staining at 1/25 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab16663 was shown to bind specifically to Cyclin D1. A band was observed at 35 kDa in wild-type A549 cell lysates with no signal observed at this size in ccnd1 knockout cell line ab286759 (knockout cell lysate ab300213). To generate this image, wild-type and ccnd1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/200 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CCND1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 33 kDa
Observed band size: 36 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab16663 observed at 36 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.
ab16663 was shown to react with CCND1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255348 (knockout cell lysate ab263808) was used. Wild-type HeLa and CCND1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab16663 and Anti-Vinculin antibody [VIN-54] overnight at 4°C at a 1 in 200 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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IHC image of ab16663 staining Cyclin D1 in a section of formalin-fixed paraffin-embedded normal human tonsil* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab16663, 1/100 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
This image was generated from the hybridoma version.
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Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/200 dilution + Wild-type HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 33 kDa
Observed band size: 36 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab16663 observed at 36 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab16663 was shown to react with Cyclin D1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261760 (knockout cell lysate ab256864) was used. Wild-type HeLa and CCND1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab16663 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 200 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence analysis of C6 (rat glial tumor glial cell) cells labeling Cyclin D1 with purified ab16663 at 1/50 (5.42µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
This image was generated from the hybridoma version.
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Intracellular flow cytometry analysis of MCF-7 (human breast carcinoma) labeling Cyclin D1 with purified ab16663 at 1/30 dilution (9.03 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlableled control - Unlabelled cells (blue). This image was generated from the hybridoma version.
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All lanes : Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/1000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate
Lane 2 : MCF7 (Human breast adenocarcinoma epithelial cell) cell lysate
Lane 3 : CCND1 KO HAP1 cell lysate
Lane 4 : HAP1 (Human chronic myelogenous leukemia near-haploid cell line) cell lysate
Lane 5 : Neuro-2a (Mouse neuroblastoma neuroblast) cell lysate
Lane 6 : NIH/3T3 (Mouse embryonic fibroblast) cell lysate
Lane 7 : C6 (Rat glial tumor glial cell) cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 33 kDa
Observed band size: 33 kDa
Exposure time: 2 seconds -
All lanes : Anti-Cyclin D1 antibody [SP4] (ab16663)
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : CCND1 (Cyclin D1) knockout HAP1 whole cell lysate
Lane 3 : A431 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 33 kDaLanes 1 - 3: Merged signal (red and green). Green - ab16663 observed at 34 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab16663 was shown to specifically recognize CCND1 (Cyclin D1) in wild-type HAP1 cells as signal was lost at the expected MW in CCND1 (Cyclin D1) knockout cells. Wild-type and CCND1 (Cyclin D1) knockout samples were subjected to SDS-PAGE. Ab16663 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/200 dilution and 1/20,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.
This image was generated from the hybridoma version.
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IHC image of ab16663 staining Cyclin D1 in rat esophagus formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16663, 1:100 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.This image was generated from the hybridoma version.
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Immunocytochemistry/ Immunofluorescence analysis of Neuro-2a (mouse neuroblastoma neuroblast) cells labeling Cyclin D1µ with purified ab16663 at 1/50 (5.42µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
This image was generated from the hybridoma version.
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ab16663 staining Cyclin D1 in MCF7 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab16663 at a working dilution of 1/250 and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594, shown in red) at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-rabbit AlexaFluor® 488 (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).This image was generated from the hybridoma version.
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Intracellular flow cytometry analysis of C6 (rat glioma) labeling Cyclin D1 with purified ab16663 at 1/30 dilution (9.03 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlableled control - Unlabelled cells (blue). This image was generated from the hybridoma version.
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Intracellular flow cytometry analysis of NIH/3T3 (mouse embryo) labeling Cyclin D1 with purified ab16663 at 1/30 dilution (9.03 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlableled control - Unlabelled cells (blue). This image was generated from the hybridoma version.
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ab16663 staining Cyclin D1 in wild-type HAP1 cells (top panel) and CCND1 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab16663 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This image was generated from the hybridoma version.
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Immunohistochemical analysis of mouse testis tissue, staining Cyclin D1 with ab16663.
Antigen retrieval was performed via Tris-EDTA buffer. Sections were blocked with 3% BSA and incubated with primary antibody (1/50) overnight at 4°C. An AlexaFluor®594-conjugated secondary antibody was used to detect staining.This image was generated from the hybridoma version.
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Anti-Cyclin D1 antibody [SP4] (ab137875) at 1/5000 dilution + MCF-7 cell lysate
Predicted band size: 33 kDaThis image was generated from the hybridoma version.
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Lane 1 : Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/200 dilution
Lane 2 : Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/400 dilution
All lanes : Whole cell lysate prepared from T24 bladder cancer cells
Lysates/proteins at 25 µg per lane.
Secondary
All lanes : Goat anti-rabbit IgG conjugated to HRP at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 33 kDa
Observed band size: 33 kDa
Exposure time: 10 minutesGel run under denaturing conditions 4-12% gradient.
This image was generated from the hybridoma version.
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Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/25 dilution + MCF7 cell lysate
Predicted band size: 33 kDa
Observed band size: 36 kDa why is the actual band size different from the predicted?This image was generated from the hybridoma version.
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ab16663 staining Cyclin D1 in Human urinary tract tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% BSA for 30 minutes at room temperature; antigen retrieval was by heat mediation in citrate buffer. Samples were incubated with primary antibody (1/100 in PBS) for 1 hour. An undiluted HRP-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.
This image was generated from the hybridoma version.
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Human mantle cell lymphoma stained with ab16663.
This image was generated from the hybridoma version.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (487)
ab16663 は 487 報の論文で使用されています。
- Wang L et al. Circ_0000520 interacts with miR-512-5p to upregulate KIAA0100 to promote malignant behaviors in lung cancer. Histol Histopathol 38:73-89 (2023). PubMed: 35866672
- Liu X et al. LncRNA ZFAS1 contributes to osteosarcoma progression via miR-520b and miR-520e-mediated inhibition of RHOC signaling. Clinics (Sao Paulo) 78:100143 (2023). PubMed: 36473367
- Chen C et al. miRNA-576-5p promotes endometrial cancer cell growth and metastasis by targeting ZBTB4. Clin Transl Oncol 25:706-720 (2023). PubMed: 36538280
- Ni T et al. PTBP1 drives c-Myc-dependent gastric cancer progression and stemness. Br J Cancer 128:1005-1018 (2023). PubMed: 36635500
- Mok KK et al. Apolipoprotein E ε4 disrupts oligodendrocyte differentiation by interfering with astrocyte-derived lipid transport. J Neurochem 165:55-75 (2023). PubMed: 36549843