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Bcl-2 Recombinant Rabbit Monoclonal Antibody [JF104-8]

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Catalog# ET1702-53

Bcl-2 Recombinant Rabbit Monoclonal Antibody [JF104-8]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Bcl-2 Recombinant Rabbit Monoclonal Antibody [JF104-8]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human Bcl-2 aa 1-230.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, FC

Molecular Weight

Predicted band size: 26 kDa

Positive Control

HeLa cell lysate, Jurkat cell lysate, MCF-7 cell lysate, HL-60 cell lysate, THP-1 cell lysate, rat spleen tissue lysate, mouse spleen tissue lysate, Hela, A549, human tonsil tissue, human colon carcinoma tissue, mouse kidney tissue, human lung carcinoma tissue, Jurkat, human B-cell lymphoma tissue.

Conjugation

unconjugated

Clone Number

JF104-8

RRID

AB_3070315

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000-1:20,000

  • IF-Cell

  • 1:100-1:200

  • IF-Tissue

  • 1:500-1:1,000

  • IHC-P

  • 1:1,000-1:5,000

  • FC

  • 1:500-1:1,000

Target

Function

Bcl-2 is one among many key regulators of apoptosis, which are essential for proper development, tissue homeostasis, and protection against foreign pathogens. Human Bcl-2 is an anti-apoptotic, membrane-associated oncoprotein that can promote cell survival through protein-protein interactions with other Bcl-2 related family members, such as the death suppressors Bcl-xL, Mcl-1, Bcl-w, and A1 or the death agonists Bax, Bak, Bik, Bad, and Bid. The anti-apoptotic function of Bcl-2 can also be regulated through proteolytic processing and phospho-rylation. Bcl-2 may promote cell survival by interfering with the activation of the cytochrome c/Apaf-1 pathway through stabilization of the mitochondrial membrane. Mutations in the Bcl-2 gene can contribute to cancers where normal physiological cell death mechanisms are compromised by deregulation of the anti-apoptotic influence of Bcl-2.

Background References

1. Cao LH et al. Morphine, a potential antagonist of cisplatin cytotoxicity, inhibits cisplatin-induced apoptosis and suppression of tumor growth in nasopharyngeal carcinoma xenografts. Sci Rep 6:18706 (2016).

2. Chen B et al. Inhibition of miR-29c promotes proliferation, and inhibits apoptosis and differentiation in P19 embryonic carcinoma cells. Mol Med Rep 13:2527-35 (2016).

Sequence Similarity

Belongs to the Bcl-2 family.

Tissue Specificity

Expressed in a variety of tissues.

Post-translational Modification

Phosphorylation/dephosphorylation on Ser-70 regulates anti-apoptotic activity. Growth factor-stimulated phosphorylation on Ser-70 by PKC is required for the anti-apoptosis activity and occurs during the G2/M phase of the cell cycle. In the absence of growth factors, BCL2 appears to be phosphorylated by other protein kinases such as ERKs and stress-activated kinases. Phosphorylated by MAPK8/JNK1 at Thr-69, Ser-70 and Ser-87, wich stimulates starvation-induced autophagy. Dephosphorylated by protein phosphatase 2A (PP2A) (By similarity).; Proteolytically cleaved by caspases during apoptosis. The cleaved protein, lacking the BH4 motif, has pro-apoptotic activity, causes the release of cytochrome c into the cytosol promoting further caspase activity.; Monoubiquitinated by PRKN, leading to increase its stability. Ubiquitinated by SCF(FBXO10), leading to its degradation by the proteasome.

Subcellular Location

Mitochondrion outer membrane, Nucleus membrane, Endoplasmic reticulum membrane, Cytoplasm.

UNIPROT

SwissProt: P10415 Human

SwissProt: P10417 Mouse

SwissProt: P49950 Rat

Synonyms

Apoptosis regulator Bcl 2 antibody

Apoptosis regulator Bcl-2 antibody

Apoptosis regulator Bcl2 antibody

AW986256 antibody

B cell CLL/lymphoma 2 antibody

B cell leukemia/lymphoma 2 antibody

Bcl-2 antibody

Bcl2 antibody

BCL2_HUMAN antibody

C430015F12Rik antibody

Expand

Images

  • Western blot analysis of Bcl-2 on different lysates with Rabbit anti-Bcl-2 antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: Jurkat cell lysate<br />Lane 3: MCF7 cell lysate<br />Lane 4: HL-60 cell lysate<br />Lane 5: THP-1 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 26 kDa<br />Observed band size: 26 kDa<br /><br />Exposure time: 1 minute;<br /><br />15% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Bcl-2 on different lysates with Rabbit anti-Bcl-2 antibody (ET1702-53) at 1/5,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: Jurkat cell lysate
    Lane 3: MCF7 cell lysate
    Lane 4: HL-60 cell lysate
    Lane 5: THP-1 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 26 kDa
    Observed band size: 26 kDa

    Exposure time: 1 minute;

    15% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-53) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Bcl-2 on different lysates with Rabbit anti-Bcl-2 antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>) at 1/5,000 dilution.<br /><br />Lane 1: Rat spleen tissue lysate<br />Lane 2: Mouse spleen tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 26 kDa<br />Observed band size: 25 kDa<br /><br />Exposure time: 2 minutes;<br /><br />12% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Bcl-2 on different lysates with Rabbit anti-Bcl-2 antibody (ET1702-53) at 1/5,000 dilution.

    Lane 1: Rat spleen tissue lysate
    Lane 2: Mouse spleen tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 26 kDa
    Observed band size: 25 kDa

    Exposure time: 2 minutes;

    12% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-53) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Bcl-2 on different lysates with Rabbit anti-Bcl-2 antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>) at 1/5,000 dilution.<br /><br />Lane 1: Hela-si NT cell lysate (10 µg/Lane)<br />Lane 2: Hela-si Bcl-2 cell lysate (10 µg/Lane)<br /><br />Predicted band size: 26 kDa<br />Observed band size: 26 kDa<br /><br />Exposure time: 31 seconds; ECL: K1801<br />4-20% SDS-PAGE gel.<br /><br /><a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a> was shown to specifically react with Bcl-2 in Hela-si NT cells. Weakened band was observed when Hela-si Bcl-2 sample was tested. Hela-si NT and Hela-si Bcl-2 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>, 1/5,000) and Loading control antibody (Rabbit anti-GAPDH, <a href="/products/ET1601-4" style="font-weight: bold;text-decoration: underline;">ET1601-4</a>, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of Bcl-2 on different lysates with Rabbit anti-Bcl-2 antibody (ET1702-53) at 1/5,000 dilution.

    Lane 1: Hela-si NT cell lysate (10 µg/Lane)
    Lane 2: Hela-si Bcl-2 cell lysate (10 µg/Lane)

    Predicted band size: 26 kDa
    Observed band size: 26 kDa

    Exposure time: 31 seconds; ECL: K1801
    4-20% SDS-PAGE gel.

    ET1702-53 was shown to specifically react with Bcl-2 in Hela-si NT cells. Weakened band was observed when Hela-si Bcl-2 sample was tested. Hela-si NT and Hela-si Bcl-2 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1702-53, 1/5,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Bcl-2 antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Bcl-2 antibody (ET1702-53) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-53) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human B-cell lymphoma tissue with Rabbit anti-Bcl-2 antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human B-cell lymphoma tissue with Rabbit anti-Bcl-2 antibody (ET1702-53) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-53) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-Bcl-2 antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-Bcl-2 antibody (ET1702-53) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-53) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Bcl-2 antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Bcl-2 antibody (ET1702-53) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-53) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-Bcl-2 antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>) at 1/4,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>) at 1/4,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-Bcl-2 antibody (ET1702-53) at 1/4,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-53) at 1/4,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of Jurkat cells labeling Bcl-2.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of Jurkat cells labeling Bcl-2.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-53, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Flow cytometric analysis of human peripheral blood lymphocytes labeling Bcl-2.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1702-53" style="font-weight: bold;text-decoration: underline;">ET1702-53</a>, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of human peripheral blood lymphocytes labeling Bcl-2.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-53, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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