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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 cellsLane 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).
Figure 1 Western Blot Validation in Human and Mouse Cell Lines
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 cellsLane B: Mouse A-20 cells
IAV
Trial Size

Bcl-2 Antibody

CATALOG NUMBER: 3335

Clonality:
Polyclonal
Tested Applications:
ELISA, IF, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
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.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IF, IHC-P, WB
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.
Positive Control 1:
Cat. No. 1288 - A20 Cell Lysate
Positive Control 2:
Cat. No. 1305 - Human Kidney Tissue Lysate
Positive Control 3:
Cat. No. 10-401 - Human Kidney Tissue Slide
Positive Control 4:
Cat. No. 1219 – MCF-7 Cell Lysate
Positive Control 5:
Cat. No. 1208 – THP-1 Cell Lysate
Predicted Molecular Weight:
Predicted: 26kD
Observed: 26 kD
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. 

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
Purification:
Bcl-2 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
Bcl-2 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
Bcl-2antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
BCL2
Additional Names:
Bcl-2 Antibody: Bcl-2, PPP1R50, Apoptosis regulator Bcl-2
Protein Accession Number:
AAH27258
Protein Gi Number:
20072668
Ncbi Gene Id Number:
596
User Note:
Optimal dilutions for each application to be determined by the researcher.
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 Reference 1:
Lockshin et al. Cell Death Differ. 2000; 7:2-7.
Background Reference 2:
Cory et al. Oncogene 2003; 22:8590-607.
Background Reference 3:
Heiser et al. Exp. Geron. 2004; 39:1125-35.
Background Reference 4:
Hockenbery et al. Nature 1990; 348:334-6.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

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.

CATALOG NUMBER:

3335

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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