Invitrogen

Cyclin A2 Monoclonal Antibody (E23.1)

3个已发表图片

6 篇参考文献

查看其他23个Cyclin A2抗体

Cyclin A2 Antibody in Immunocytochemistry (ICC/IF)

Cyclin A2 Antibody in Immunocytochemistry (ICC/IF)

图: 1 / 14

Cyclin A2 Antibody (MA1-154) in ICC/IF

Immunofluorescence analysis of Cyclin A2 was performed using 70% confluent log phase HeLa cells treated with Camptothecin (100nM, 16 hrs). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Cyclin A2 Monoclonal Antibody (Product # MA1-154) at 1:50 dilution in 0... View More

Cyclin A2 Antibody in Immunocytochemistry (ICC/IF)

Cyclin A2 Antibody in Immunocytochemistry (ICC/IF)

Cyclin A2 Antibody in Immunohistochemistry (Paraffin) (IHC (P))

Cyclin A2 Antibody in Western Blot (WB)

Cyclin A2 Antibody in Western Blot (WB)

Cyclin A2 Antibody in Western Blot (WB)

Cyclin A2 Antibody in Western Blot (WB)

Cyclin A2 Antibody in Immunoprecipitation (IP)

Cyclin A2 Antibody in Western Blot (WB)

Cyclin A2 Antibody in Western Blot (WB)

Cyclin A2 Antibody in Western Blot (WB)

Cyclin A2 Antibody

Cyclin A2 Antibody

Cyclin A2 Antibody

Product Image Gallery

Cyclin A2 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Cyclin A2 was performed using 70% confluent log phase HeLa cells treated with Camptothecin (100nM, 16 hrs). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Cyclin A2 Monoclonal Antibody (Product # MA1-154) at 1:50 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing accumulation of Cyclin A2 in the nucleus upon treatment. Panel e represents the control cells showing cytoplasmic staining. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Cyclin A2 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Cyclin A (green) in HeLa cells left untreated (UT, left panels) or treated with 100nM camptothecin (CPT, right panels). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a Cyclin A monoclonal antibody (Product # MA1-154) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502). Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249; lower panels) for 30 minutes. Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification. Note: Red arrows indicate nuclear translocation of Cyclin A in response to treatment.

Cyclin A2 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of Cyclin A showing staining in the nucleus of paraffin-embedded human tonsil tissue (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Cyclin A monoclonal antibody (Product # MA1-154) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Cyclin A2 Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extract (30 µg lysate) of HeLa (Lane 1), HeLa treated with doxorubicin (0.5uM, 24 hrs) (Lane 2), COS-7 (Lane 3), A431 (Lane 4) and Jeko1(Lane 5). The blot was probed with Anti-Cyclin A2 Monoclonal Antibody (E23.1) (Product # MA1-154, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). A 52 kDa band corresponding to Cyclin A2 was observed in all cell the lines tested and was increased upon Doxorubicin treatment.

Cyclin A2 Antibody in Western Blot (WB) — Western blot analysis of Cyclin A was performed by loading 10 µg of COS7 cell lysates from cells left untreated (UT), or treated with 2mM hydroxyurea (HU), or 100 nM camptothecin (CPT) for 16 hours per lane onto a 4-12% Bis-Tris polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% BSA in TBST for at least 1 hour. The membrane was probed with a Cyclin A monoclonal antibody (Product # MA1-154) at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBST, and probed with an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) at a dilution of 1:20,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).

Cyclin A2 Antibody in Western Blot (WB) — Western blot analysis of Cyclin A was performed by loading 10 µg of lysates from HEK293T cells left untreated (UT), or treated with 2mM hydroxyurea (HU) or 100 nM camptothecin (CPT) for 16 hours onto a 4-12% Bis-Tris polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% BSA in TBST for at least 1 hour. Membranes were probed with a Cyclin A monoclonal antibody (Product # MA1-154) at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBST, and probed with an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) at a dilution of 1:20,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).

Cyclin A2 Antibody in Western Blot (WB) — Western blot analysis of Cyclin A was performed by loading 10 µg of lysates from HeLa cells left untreated (UT), or treated with 2mM hydroxyurea (HU) or 100 nM camptothecin (CPT) for 16 hours onto a 4-12% Bis-Tris polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% BSA in TBST for at least 1 hour. Membranes were probed with a Cyclin A monoclonal antibody (Product # MA1-154) at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBST, and probed with an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) at a dilution of 1:20,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).

Cyclin A2 Antibody in Immunoprecipitation (IP) — Immunoprecipitation analysis of Cyclin A was performed on HeLa cell lysates, from cells left untreated (UT) or treated with 100 nM camptothecin (CPT). Antigen-antibody complexes were formed by incubating 375 µg of lysate with 1 µg of a Cyclin A monoclonal antibody (Product # MA1-154) overnight on an end-over-end rotator at 4°C. The immune complexes were captured on 100 µL Protein A/G Plus Agarose (Product # 20423), washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer (Product # 39000). Samples, including control input cell lysates (-IP, left 2 lanes), were resolved on a 4-12% Bis-Tris polyacrylamide gel, transferred to a nitrocellulose membrane and blocked with 5% BSA/TBST for 1 hour. The membrane was probed with a Cyclin A monoclonal antibody (Product # MA1-154) at a dilution of 1:1000 overnight rotating at 4°C, washed in TBST, and probed with Clean-Blot IP Detection Reagent (Product # 21230). Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).

Cyclin A2 Antibody in Western Blot (WB) — Ki16425 decreases lysophosphatidic acid (LPA)-induced expression of cyclin A2 and D1, CDK2, and CDK4 in HaCaT cells. HaCaT cells were seeded in 60-mm dishes and starved with serum-free media containing 0.1% fatty-acid-free bovine serum albumin (FAF-BSA) for 12 to 16 h. The cells were treated with 10 muM LPA in the presence or absence of 10 muM ki16425 for 6 h. Cyclin A2, D1, and E2, CDK2, CDK4, and p27 Kip1 protein levels were analyzed by Western blotting, quantified using ImageJ software, and normalized to those of beta-actin. Representative images of the blots ( A , C ) and relative densitometric bar graphs ( B , D ) are shown. The data are presented as the mean +- standard error of the mean (SEM) of results obtained from three to four independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.005 vs. Con; # p < 0.05, ## p < 0.01 vs. LPA.

Cyclin A2 Antibody in Western Blot (WB) — Figure 5 LPAR1 mediates the lysophosphatidic acid (LPA)-induced HaCaT cell proliferation. ( A ) HaCaT cells were seeded in six-well plates and treated with cytokine mixture (CM; 20 ng/mL tumor necrosis factor (TNF)-alpha and 10 ng/mL interleukin (IL)-6) for 4 h. LPAR1, LPAR2, and LPAR3 mRNA expression was analyzed by qRT-PCR. ( B ) HaCaT cells were seeded in four-well chambers and treated with CM for 24 h. Immunocytochemical analysis was performed using an anti-LPAR1 antibody and an Alexa Fluor 488-conjugated secondary antibody. The nuclei were counterstained with DAPI (blue). Representative staining images (left panel) and percentage of LPAR1 positive cells were quantified using confocal microscope (original magnification, 400x; the scale bars represent 20 mum). ( C - E ) HaCaT cells were transfected with a control siRNA (siCon) or LPAR1 siRNA (siLPAR1) for 6 h and treated with LPA for 24 h. ( C ) The mRNA expression of LPAR1 was analyzed by qRT-PCR. ( D ) Cell viability was analyzed by CCK8 assay and calculated as a percentage relative to that of siCon-BSA cells. ( E ) The cell morphology was observed by light microscopy. Original magnification, 100x. ( F , G ) HaCaT cells were transfected with siCon or siLPAR1 for 6 h and treated with LPA for 6 h. The levels of the proteins LPAR1, cyclin A2, CDK2, and p27 Kip1 were analyzed by Western blotting, quantified using ImageJ software, and normalized to those of beta-actin. (F) Representative images of the blots and ( G ) relative

Cyclin A2 Antibody in Western Blot (WB) — Figure 3 Effect of survivin inhibition (A) Survivin silencing by siRNA on RC-4 B/C cells. (B) Survivin inhibition by YM155, a small molecule inhibitor in RC-4 B/C cells. The effect of YM155 (C) and survivin siRNA (D) on pituitary adenoma cell viability, proliferation and cell cycle phases of RC-4 B/C cells (see details in the text). (E) Decrease of CCNA2, CDK2 mRNA and CCNA2, total CDK2 and p-CDK2 protein abundance following ASA treatment in RC-4 B/C cells. Left: mRNA, middle and right : western blot and densitometry. (F) YM155 and survivin siRNA transfection did not significantly alter CCNA2 and CDK2 expression in RC-4 B/C cells. (G) Survivin overexpression did not reduce the viability of RC-4 B/C cells. * : p<0.05; ** : p<0.01; *** : p<0.001; **** : p<0.0001.

Cyclin A2 Antibody — Altered expression of target protein upon cell treatment demonstrates antibody specificity. Immunofluorescence of Cyclin A2 using with Cyclin A2 Monoclonal Antibody (E23.1) (Product # MA1-154) shows an increase and translocation to the nucleus upon treatment with Camptothecin. {TM}

Cyclin A2 Antibody — Altered expression of target protein upon cell treatment demonstrates antibody specificity. Western blot analysis of Cyclin A2 using a Cyclin A2 Monoclonal Antibody (E23.1) (Product # MA1-154) shows an induction upon doxorubicin treatment. {TM}

Cyclin A2 Antibody — Figure 3 Effect of survivin inhibition (A) Survivin silencing by siRNA on RC-4 B/C cells. (B) Survivin inhibition by YM155, a small molecule inhibitor in RC-4 B/C cells. The effect of YM155 (C) and survivin siRNA (D) on pituitary adenoma cell viability, proliferation and cell cycle phases of RC-4 B/C cells (see details in the text). (E) Decrease of CCNA2, CDK2 mRNA and CCNA2, total CDK2 and p-CDK2 protein abundance following ASA treatment in RC-4 B/C cells. Left: mRNA, middle and right : western blot and densitometry. (F) YM155 and survivin siRNA transfection did not significantly alter CCNA2 and CDK2 expression in RC-4 B/C cells. (G) Survivin overexpression did not reduce the viability of RC-4 B/C cells. * : p<0.05; ** : p<0.01; *** : p<0.001; **** : p<0.0001. {TM}

产品信息

MA1-154

应用

建议稀释比

已发表文章

免疫印迹 (WB)

1:1,000

查看6篇已发表文章 6篇已发表文章

免疫组化（石蜡） (IHC (P))

1:50-1:500

-

免疫组化（冰冻） (IHC (F))

Assay-dependent

-

免疫细胞化学 (ICC/IF)

1:50

-

免疫沉淀 (IP)

1 µg

-

产品规格

种属反应

Bovine, Human, Mouse, Non-human primate, Rat

已发表种属

Human, Rat

宿主/亚型

Mouse / IgG2a

分类

Monoclonal

类型

Antibody

克隆号

E23.1

抗原

Bovine Cyclin A fusion protein

偶联物

Unconjugated Unconjugated Unconjugated

形式

Liquid

浓度

1 mg/mL

纯化类型

Protein A

保存液

PBS with 1mg/mL BSA, 30% glycerol

内含物

0.05% sodium azide

保存条件

-20°C

运输条件

Ambient (domestic); Wet ice (international)

AB_2536862

产品详细信息

MA1-154 detects Cyclin A in mammalian samples and has been successfully used in Immunohistochemistry (Frozen and Paraffin), Immunofluorescence, Immunoprecipitation and Western Blot procedures. Immunoprecipitation and Western Blot analysis with MA1-154 show the accumulation of a prominent band at ~50 kDa in camptothecin and hydroxyurea treated cells. In Immunofluorescence applications, MA1-154 shows nuclear translocation of Cyclin A in G2/M arrested cells as a result of camptothecin treatment.

靶标信息

Cyclins are regulatory subunits of the cyclin-dependent kinases (cdks) and they control transition at different specific phases of the cell cycle. The temporal expression of cyclins is tightly regulated and subsequently plays a critical role in controlling the enzymatic activity of cdks. These cyclin / cdk complexes are essential for passage through specific stages in the cell cycle. In mammalian somatic cells, cyclin A is required for S phase and passage through G2-phase.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

Product References

Lysophosphatidic Acid Mediates Imiquimod-Induced Psoriasis-like Symptoms by Promoting Keratinocyte Proliferation through LPAR1/ROCK2/PI3K/AKT Signaling Pathway.

International journal of molecular sciences

Kim D,Kim HJ,Baek JO,Roh JY,Jun HS

Published figure using Cyclin A2 monoclonal antibody (Product # MA1-154) in Western Blot

Tue Oct 05 00:00:00 UTC 2021

Single-tubule RNA-Seq uncovers signaling mechanisms that defend against hyponatremia in SIADH.

Kidney international

Lee JW,Alsady M,Chou CL,de Groot T,Deen PMT,Knepper MA,Ecelbarger CM

MA1-154 was used in Western Blotting to identify signalling pathways activated at the onset of vasopressin-escape.

Mon Jan 01 00:00:00 UTC 2018

Survivin as a potential therapeutic target of acetylsalicylic acid in pituitary adenomas.

Oncotarget

Németh K,Szücs N,Czirják S,Reiniger L,Szabó B,Barna G,Karászi K,Igaz P,Zivkovic V,Korbonits M,Patócs A,Butz H

Published figure using Cyclin A2 monoclonal antibody (Product # MA1-154) in Western Blot

Tue Jun 26 00:00:00 UTC 2018

APOLLON protein promotes early mitotic CYCLIN A degradation independent of the spindle assembly checkpoint.

The Journal of biological chemistry

Kikuchi R,Ohata H,Ohoka N,Kawabata A,Naito M

MA1-154 was used in western blot to study the spindle assembly checkpoint-independent regulation of mitotic cyclin A by the APOLLON protein

Fri Feb 07 00:00:00 UTC 2014

Disrupting Skp2-cyclin A interaction with a blocking peptide induces selective cancer cell killing.

Molecular cancer therapeutics

Ji P,Sun D,Wang H,Bauzon F,Zhu L

MA1-154 was used in immunoprecipitation and western blot to report that the Skp2-cyclin A interaction has a major role in cancer cell survival

Thu Feb 01 00:00:00 UTC 2007

Differential apoptosis by gallotannin in human colon cancer cells with distinct p53 status.

Molecular carcinogenesis

Al-Ayyoubi S,Gali-Muhtasib H

MA1-154 was used in western blot to elucidate how treatment with gallotannin prevents cancer

Thu Mar 01 00:00:00 UTC 2007

生物信息学

蛋白别名: Cyclin A; Cyclin A2 (Cyclin A); Cyclin-A; Cyclin-A2

查看更多收起

基因别名: AA408589; Ccn-1; CCN1; CCNA; CCNA2; Cyca; CycA2

查看更多收起

UniProt ID: (Bovine) P30274, (Human) P20248, (Mouse) P51943

查看更多收起

Entrez Gene ID: (Bovine) 281667, (Human) 890, (Rat) 114494, (Mouse) 12428

查看更多收起

protein binding protein kinase binding molecular_function kinase activator

G2/M transition of mitotic cell cycle regulation of DNA replication cell cycle G1/S phase transition cell division regulation of cyclin-dependent protein serine/threonine kinase activity mitotic nuclear division mitotic G2 DNA damage checkpoint Ras protein signal transduction viral process organ regeneration response to glucagon cellular response to platelet-derived growth factor stimulus cellular response to leptin stimulus positive regulation of transcription, DNA-templated positive regulation of fibroblast proliferation cellular response to cocaine cellular response to luteinizing hormone stimulus cellular response to estradiol stimulus cellular response to hypoxia cellular response to nitric oxide cochlea development cellular response to insulin-like growth factor stimulus regulation of G2/M transition of mitotic cell cycle response to estradiol cell cycle