Bcl-2 Antibody

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ProSci
Product Code: PSI-3335
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-3335-0.02mg0.02mg£150.00
Quantity:
PSI-3335-0.1mg0.1mg£449.00
Quantity:
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Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunofluorescence (IF)
  • Immunohistochemistry (IHC)
  • Western Blot (WB)

Images

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<strong>Figure 1 Western Blot Validation in Human and Mouse Cell Lines </strong><br> Loading: 15 μg of lysates per lane. Antibodies: Bcl-2 3335, (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane A: Human Daudi cells Lane B: Mouse A-20 cells
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<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: Bcl-2 3335, (2 μg/mL), Bcl-2 3337, (2 μg/mL), beta-actin (1 μg/mL) and GAPDH (0.02 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 3 Western Blot Validation in Human Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: Bcl-2 3335, (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 4 Immunohistochemistry Validation of Bcl-2 in Human Kidney </strong><br> Immunohistochemical analysis of paraffin-embedded Human Kidney using anti-Bcl-2 antibody (3335) at 2μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
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<strong>Figure 5 Immunofluorescence Validation of Bcl-2 in Human Kidney Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed Human Kidney Cells labeling Bcl-2 with 3335 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
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<strong>Figure 6 Immunohistochemistry Validation of Bcl-2 in Mouse Brain Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded Mouse Brain Tissue using anti-Bcl-2 antibody (3335) at 5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
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<strong>Figure 7 Immunofluorescence Validation of Bcl-2 in Mouse Brain Tissue</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed Mouse Brain Tissue labeling Bcl-2 with 3335 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green).
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<strong>Figure 1 Western Blot Validation in Human and Mouse Cell Lines </strong><br> Loading: 15 μg of lysates per lane. Antibodies: Bcl-2 3335, (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane A: Human Daudi cells Lane B: Mouse A-20 cells
<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: Bcl-2 3335, (2 μg/mL), Bcl-2 3337, (2 μg/mL), beta-actin (1 μg/mL) and GAPDH (0.02 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Western Blot Validation in Human Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: Bcl-2 3335, (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Immunohistochemistry Validation of Bcl-2 in Human Kidney </strong><br> Immunohistochemical analysis of paraffin-embedded Human Kidney using anti-Bcl-2 antibody (3335) at 2μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 5 Immunofluorescence Validation of Bcl-2 in Human Kidney Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed Human Kidney Cells labeling Bcl-2 with 3335 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
<strong>Figure 6 Immunohistochemistry Validation of Bcl-2 in Mouse Brain Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded Mouse Brain Tissue using anti-Bcl-2 antibody (3335) at 5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 7 Immunofluorescence Validation of Bcl-2 in Mouse Brain Tissue</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed Mouse Brain Tissue labeling Bcl-2 with 3335 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green).

Documents

Further Information

Additional Names:
Bcl-2 Antibody: Bcl-2, PPP1R50, Apoptosis regulator Bcl-2
Application Note:
WB: 1-2μg/mL; IHC-P: 2 μg/mL; IF: 10 μg/mL.

Antibody validated: Western Blot in human and mouse samples; Immunohistochemistry in human and mouse samples ; Immunofluorescence in human and mouse samples. All other applications and species not yet tested.
Background:
Bcl-2 Antibody: Apoptosis plays a major role in normal organism development, tissue homeostasis, and removal of damaged cells. Disruption of this process has been implicated in a variety of diseases such as cancer. Bcl-2 is the founding member of a family of over 20 proteins that are critical regulators of apoptosis. These can be divided into two classes: those that inhibit apoptosis and those that promote cell death. Bcl-2 is an inner mitochondrial membrane protein that inhibits apoptosis. It is thought to act by interacting with pro-apoptotic Bcl-2 family members such as Bak and Bad. Overexpression of Bcl-2 has been linked to human cancers such as B-cell lymphoma and prostate cancer.
Background References:
  • Lockshin et al. Cell Death Differ. 2000; 7:2-7.
  • Cory et al. Oncogene 2003; 22:8590-607.
  • Heiser et al. Exp. Geron. 2004; 39:1125-35.
  • Hockenbery et al. Nature 1990; 348:334-6.
Buffer:
Bcl-2 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Conjugate:
Unconjugated
DISCLAIMER:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.
Homology:
Predicted species reactivity based on immunogen sequence: Bovine: (100%), Rat: (100%), Chicken: (93%)
Immunogen:
Anti-Bcl-2 antibody (3335) was raised against a peptide corresponding to 15 amino acids near the amino terminus of human Bcl-2.

The immunogen is located within the first 50 amino acids of Bcl-2.
ISOFORMS:
Human Bcl-2 has 2 isoforms, including isoform 1 (239aa, 26kD), and isoform 2 (205aa, 22kD). Mouse Bcl-2 has 2 isoforms, including isoform 1 (236aa, 26kD), and isoform 2 (199aa, 22kD). Rat Bcl-2 has one isoform (236aa, 27kD). 3337 can detect all isoforms of human, mouse and rat
NCBI Gene ID #:
596
NCBI Official Name:
B-cell CLL/lymphoma 2
NCBI Official Symbol:
BCL2
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 26kD

Observed: 26 kD
Protein Accession #:
AAH27258
Protein GI Number:
20072668
Purification:
Bcl-2 Antibody is affinity chromatography purified via peptide column.
Research Area:
Apoptosis,Autophagy,Cancer
Swissprot #:
P10415
User NOte:
Optimal dilutions for each application to be determined by the researcher.
VALIDATION:

Independent Antibody Validation in Cell lines (Figure 2) shows similar Bcl-2 expression profile in human and mouse cell lines detected by two independent anti-Bcl-2 antibodies that recognize different epitopes, 3335 against N-terminus domain and 3337 against central domain.  Bcl-2 proteins are detected in several tested cell lines at different expression levels by the two independent antibodies. 

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