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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [6]
- Immunocytochemistry [1]
- Immunohistochemistry [2]
- Flow cytometry [1]
- Other assay [1]
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- Product number
- MA5-17052 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CDK2 Monoclonal Antibody (1A6)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- MA5-17052 targets CDK2 in FACS, ICC, IHC, IF and WB applications and shows reactivity with Human and Mouse samples.
- Antibody clone number
- 1A6
- Concentration
- 1 mg/mL
Submitted references MM-129 as a Novel Inhibitor Targeting PI3K/AKT/mTOR and PD-L1 in Colorectal Cancer.
Hermanowicz JM, Pawlak K, Sieklucka B, Czarnomysy R, Kwiatkowska I, Kazberuk A, Surazynski A, Mojzych M, Pawlak D
Cancers 2021 Jun 26;13(13)
Cancers 2021 Jun 26;13(13)
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of CDK2 using a CDK2 monoclonal antibody (Product # MA5-17052) against a human CDK2 (AA: 197-295) recombinant protein.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of CDK2 using CDK2 monoclonal antibody (Product # MA5-17052) in Jurkat (1), HL-60 (2), K562 (3), A431 (4), HeLa (5), and NIH3T3 (6) cell lysate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of A-431 (Lane 1), U-87 MG (Lane 2), Jurkat (Lane 3), MCF7 (Lane 4), K-562 (Lane 5), NIH/3T3 (Lane 6), Caki-1 (Lane 7) and Hep G2 (Lane 8). The blot was probed with Anti-CDK2 Monoclonal Antibody (Product # MA5-17052, 1:2000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25µg/ml, 1:4000 dilution). A 34 kDa band corresponding to CDK2 was observed across the cell lines tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of CDK2 using a CDK2 monoclonal antibody (Product # MA5-17052) against a human CDK2 (AA: 197-295) recombinant protein.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of CDK2 using CDK2 monoclonal antibody (Product # MA5-17052) in Jurkat (1), HL-60 (2), K562 (3), A431 (4), HeLa (5), and NIH3T3 (6) cell lysate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockout of CDK2 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR692361_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of CDK2 was performed by loading 30 µg of A-431 wild type (Lane 1), A-431 Cas9 (Lane 2) andA-431 CDK2 KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-CDK2 Monoclonal Antibody (1A6) (Product # MA5-17052, 1:2000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:5000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to CDK2. An uncharacterized band was observed at ~55 kDa in all the samples.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of HeLa cells using CDK2 monoclonal antibody (Product # MA5-17052) (Green). Red: actin filaments have been labeled with phalloidin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded cervical cancer tissues using CDK2 monoclonal antibody (Product # MA5-17052) followed with DAB staining.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded colon cancer tissues using CDK2 monoclonal antibody (Product # MA5-17052) followed with DAB staining.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometric analysis of Jurkat cells using CDK2 monoclonal antibody (Product # MA5-17052) (green) and negative control (red).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 8 Phosphorylated CDK2 (p-CDK2), total CDK2 (t-CDK2), and beta-actin expression as determined by Western blot ( a ) and phosphorylated CDK2 (p-CDK2) determined by confocal microscopy ( d ) in DLD-1 and HT-29 cells treated with 5-FU (5-FU 50 uM), MM-129 (MM 1 uM, 10 uM, 100 uM), and their combination (MM 10 uM + 5-FU 50 uM) for 24 h. The samples used for electrophoresis consisted of 20 ug of protein from 6 pooled cell extracts from independent experiments ( n = 6). Band staining was quantified by densitometry ( b , c ). The corresponding uncropped blots are shown in Supplementary Figures S9a-c and S10a-c . Cells were incubated with rabbit polyclonal antibody against phospho CDK2 and secondary goat polyclonal antibody against rabbit (red label). The nuclei were stained with Hoechst 33342 (blue label) ( d ). The results are presented as means +- SDs. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. CON, ^^ p < 0.01 vs. 5-FU, ^^^ p < 0.001 vs. 5-FU, ### p < 0.001 vs. MM-129 at dose 10 uM.
