- PA5-16777 - Invitrogen Antibodies CCND1 antibody

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Molecular cell 2021 Feb 18;81(4):708-723.e5

Downregulation of ROS1 enhances the therapeutic efficacy of arsenic trioxide in acute myeloid leukemia cell lines.

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Oncology letters 2018 Jun;15(6):9392-9396

Aberrant Activation of the RANK Signaling Receptor Induces Murine Salivary Gland Tumors.

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PloS one 2015;10(6):e0128467

Chronic Deletion and Acute Knockdown of Parkin Have Differential Responses to Acetaminophen-induced Mitophagy and Liver Injury in Mice.

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The Journal of biological chemistry 2015 Apr 24;290(17):10934-46

Synergistic signaling of KRAS and thyroid hormone receptor β mutants promotes undifferentiated thyroid cancer through MYC up-regulation.

Zhu X, Zhao L, Park JW, Willingham MC, Cheng SY
Neoplasia (New York, N.Y.) 2014 Sep;16(9):757-69

Progesterone receptor activation downregulates GATA3 by transcriptional repression and increased protein turnover promoting breast tumor growth.

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Breast cancer research : BCR 2014 Dec 6;16(6):491

Smoothened controls cyclin D2 expression and regulates the generation of intermediate progenitors in the developing cortex.

Komada M, Iguchi T, Takeda T, Ishibashi M, Sato M
Neuroscience letters 2013 Jun 28;547:87-91

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Epithelial progesterone receptor exhibits pleiotropic roles in uterine development and function.

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FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2012 Mar;26(3):1218-27

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed using whole cell extracts (30 µg) of U-87 MG (Lane 1), Hep G2 (Lane 2) and SH-SY5Y (Lane 3). The blots was probed with Anti-Cyclin D1/ Bcl-1 Rabbit Polyclonal Antibody (Product # PA5-16777, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4µg/mL, 1:2500 dilution). A ~ 34 kDa band corresponding to Cyclin D1/ Bcl-1 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE®12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product # SLF2000S). Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of CCND1 was performed with 10 µg of A549 cells transfected with Transfection Reagent alone (Lane 1), 100nM Non-Targeting control siRNA (Lane 2), or 100nM siRNA against CCND1 (Lane 3). Proteins were resolved using a NuPAGE® Novex 4-12% Bis-Tris Gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002), and a protein size ladder. Proteins were wet transferred to a Pierce Nitrocellulose Membrane (Product # 88025) OR Pierce PVDF Membrane (Product # 88518) and blocked with Pierce Starting Block T20 (PBS) Blocking Buffer (Product # 37539) for 1 hour at room temperature. CCND1 was detected at ~ 36 kDa using CCND1 Rabbit polyclonal antibody (Product # PA5-16777) diluted in Pierce Starting Block T20 (PBS) Blocking Buffer 4°C overnight on a rocking platform. Pierce Goat Anti-Rabbit (Product # 31461) HRP-Conjugated Antibodies at a 1:2500 dilution were used and chemiluminescent detection was performed using Pierce Supersignal West Dura Maximum Sensitivity Substrate (Product # 37071). Relative density of the bands normalized to Lamin A/C (74/65 kDa). CCND1 Antibody (Product # PA5-16777) confirms silencing of CCND1 expression.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Formalin-fixed, paraffin-embedded human mantle cell lymphoma stained with Cyclin D1 antibody using peroxidase-conjugate and DAB. Note nuclear staining of tumor cells

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Cyclin D1/Bcl-1 showing staining in the nucleus of paraffin-embedded human breast carcinoma (right) compared to 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 D1/Bcl-1 Rabbit Polyclonal Antibody (Product # PA5-16777) diluted in 3% BSA-PBS at a dilution of 1:100 for 1 hour at 37°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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Cyclin D1/Bcl-1 showing staining in the nucleus and weak cytoplasm of paraffin-embedded mouse liver tissue (right) compared to 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 D1/Bcl-1 Rabbit Polyclonal Antibody (Product # PA5-16777) diluted in 3% BSA-PBS at a dilution of 1:20 for 1 hour at 37°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.

PA5-16777

Antibody data