BMS1028

antibody from Invitrogen Antibodies

Targeting: BCL2 Bcl-2, PPP1R50

ELISA

Immunohistochemistry

Flow cytometry

Antibody data

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Validation data

Product number: BMS1028 - Provider product page
Provider: Invitrogen Antibodies
Product name: Bcl-2 Monoclonal Antibody (Bcl-2/100), eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: Bcl-2 is a key regulator of apoptosis and the founding member of Bcl-2 family of apoptosis regulatory protiens. The anti-apoptotic functions of Bcl-2 is achieved by activation or inactivation of an inner mitochondrial permeability transition pore, which is involved in the regulation of matrix Ca2+, pH, and voltage. Applications Tested: ELISA, Flow Cytometry, Immunohistochemistry, Western Blotting.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: Bcl-2/100
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: -20°C

BCL2 protein structure - BMS1028 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of MCF-7 (Lane 1), Raji (Lane 2), T-47D (Lane 3), MDA-MB-231 (Lane 4), HeLa (Lane 5) and Jurkat (Lane 6). The blot was probed with Anti-Bcl-2 Monoclonal Antibody (Product # BMS1028, 1:500 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 27 kDa band corresponding to Bcl-2 was observed across the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Bcl-2 was achieved by transfecting HeLa cells with Bcl-2 specific validated siRNAs (Silencer® select Product # s194310, s1915). Western blot analysis (Fig. a) was performed using whole cell extracts from the Bcl-2 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Bcl-2 Monoclonal Antibody (Product # BMS1028, 1:500 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Bcl-2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of MCF-7 (Lane 1), Raji (Lane 2), T-47D (Lane 3), MDA-MB-231 (Lane 4), HeLa (Lane 5) and Jurkat (Lane 6). The blot was probed with Anti-Bcl-2 Monoclonal Antibody (Product # BMS1028, 1:500 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 27 kDa band corresponding to Bcl-2 was observed across the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of BCL2 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR946648_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of BCL2 was performed by loading 30 µg of HeLa wild type (Lane 1), HeLa Cas9 (Lane 2) and Hela BCL2 KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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-Bcl-2 Monoclonal Antibody (Bcl-2/100), eBioscience™ (Product # BMS1028, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to BCL2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1. 2-ME combined with AA promotes CML cell apoptosis. K562 and KCL22 cells were treated with 2-ME and AA. (a) Viability of cells treated with different concentrations of 2-ME and AA for 48 h was measured using MTT assay, and IC50 was calculated. IC50 doses of 2-ME and AA were used for subsequent cell treatment. (b) Cell proliferation activity at different time points examined using MTT assay. (c) Cell apoptosis measured using flow cytometry. (d) Levels of Bcl-2 and Caspase 3 measured using Western blotting. Cell experiment was conducted 3 times independently. Data were described as mean +- standard deviation. Data were analyzed using one-way ANOVA, followed by Tukey's multiple comparison test, ** p < 0.01 vs . DMSO group.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3. 2-ME combined with AA upregulates intracellular miR-223 expression. 2-ME + AA treated K562 and KCL22 cells were delivered with miR-223 inhibitor, with inhibitor NC as control. (a) miR-223 expression tested using RT-qPCR. (b) Cell proliferation at different times measured using MTT assay. (c) Cell apoptosis measured using flow cytometry. (d) Protein levels of Bcl-2 and Caspase 3 measured using Western blotting. (e) Content of ROS. F: Change of MMP. Cell experiment was conducted 3 times independently. Data were described as mean +- standard deviation. Data were analyzed using one-way ANOVA, followed by Tukey's multiple comparison test, *** p < 0.001. inhi-NC: 2-ME + AA + inhibitor NC; inhibitor: 2-ME + AA + miR-223 inhibitor.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5. Upregulation of FLT3 partially reverses the promoting effect of 2-ME + AA on apoptosis of K562 cells. 2-ME + AA treated K562 cells were delivered with pcDNA3.1-FLT3, with pcDNA3.1-NC as control. (a) FLT3 expression tested using RT-qPCR. (b) Cell proliferation at different times measured using MTT assay. (c) Cell apoptosis measured using flow cytometry. (d) Protein levels of Bcl-2 and Caspase 3 examined using Western blotting. (e) Content of ROS. (f) Change of MMP. Cell experiment was conducted 3 times independently. Data were described as mean +- standard deviation. Data were analyzed using one-way ANOVA, followed by Tukey's multiple comparison test, *** p < 0.001. pc-NC: 2-ME + AA + pcDNA3.1-NC; pc-FLT3: 2-ME + AA + pcDNA3.1-FLT3.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7. 2-ME combined with AA inhibited CML xenograft growth in mice. CML xenograft nude mouse models were established, and then intravenously injected with 2-ME, AA alone, or a combination of 2-ME and AA. (a) Image of mouse tumors. (b) Average tumor volume. (c) Average tumor weight. (d) Protein levels of Bcl-2 and Caspase 3 examined using Western blotting. N = 8, data were presented as mean +- standard deviation. Data among multiple groups were analyzed using one-way ANOVA, followed by Tukey's multiple comparison test, ** p < 0.01.