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Human Bcl-2 Minus C-Terminus Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB827

R&D Systems, part of Bio-Techne
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MAB827
MAB827-SP

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Bovine

Applications

Validated:

Simple Western, Western Blot

Cited:

ELISA-Based Protein Pin Array, Immunocytochemistry, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 118701

Product Specifications

Immunogen

E. coli-derived recombinant human Bcl-2 minus the carboxyl-terminal mitochondrial targeting sequence
Met1-Asp211
Accession # P10415

Specificity

Detects human Bcl-2 in Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human Bcl-2 Minus C-Terminus Antibody

Detection of Human Bcl-2 Minus C-Terminus antibody by Western Blot.

Detection of Human Bcl‑2 Minus C-Terminus by Western Blot.

Western blot shows lysates of KG-1 human acute myelogenous leukemia cell line. PVDF membrane was probed with 0.1-0.5 µg/mL of Mouse Anti-Human Bcl-2 Minus C-Terminus Monoclonal Antibody (Catalog # MAB827) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Bcl-2 Minus C-Terminus at approximately 24 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Bcl-2 antibody by Simple WesternTM.

Detection of Human Bcl‑2 by Simple WesternTM.

Simple Western lane view shows lysates of MCF‑7 human breast cancer cell line, loaded at 0.5 mg/mL. A specific band was detected for Bcl‑2 at approximately 24 kDa (as indicated) using 5 µg/mL of Mouse Anti-Human Bcl‑2 Minus C-Terminus Monoclonal Antibody (Catalog # MAB827). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of Human Bcl-2 by Simple Western

Detection of Human Bcl-2 by Simple Western

THZ1, suppresses Mcl-1 at the transcriptional level by disruption of its super-enhancer. (a) Real-time PCR analysis of Mcl1 mRNA levels in several GBM cells treated with vehicle or THZ1 for 24 h (n = 4). U87, U251, LN229, KNS42, and GBM22 cells were treated with 100 nM THZ1 while GBM14 was treated with 50 nM THZ1. Statistical significance was determined by two-tailed Student’s t-test; (b) U87 GBM cells were treated with DMSO or THZ1 100 nM for 24 h, subjected to CHIP with H3K27ac antibody and submitted for next generation sequencing. Super-enhancers were called using HOMER and depicted as a heatmap. The middle of each plot highlights the center of the super-enhancers (from −50 kb to 50 kb). The super enhancers are ranked by size and intensity levels are provided in the legend. The scale bar indicates the intensities. Blue depicts a high intensity level and red depicts a low intensity level; (c) A representation of global disruption of the super-enhancer landscape of U87 treated with DMSO or THZ1 100 nM in (b); (d) Shown are CHIP-seq (H3K27ac) tracks around the MCL1 locus (pile up values are indicated) in U87 treated with DMSO or THZ1 100 nM (super enhancer related to the Mcl-1 gene: chr1:150,601,879-150,630,909 (GRCh38/hg38)); (e) Standard western blots of cell lysates of U87 and GBM22 cells treated with increasing concentration of THZ1 for 24 h (pRpb1 corresponds to serine 5). Actin is used as a loading control. The protein expression levels were quantified using ImageJ (shown in cursive font); (f) Standard western blots or protein capillary electrophoresis of cell lysates of U87, U251, LN229, and GBM22 cells treated with increasing concentration of THZ1 for 24 h. Actin is used as a loading control in standard western blots and Vinculin is used as a loading control in protein capillary electrophoresis. The protein expression levels were quantified by using ImageJ (shown in cursive font). Uncropped blots are shown in Figure S10; (g) Shown are the protein expression levels of Mcl1, Noxa, Bcl2, and Bcl-xL following treatment with increasing concentration of THZ1 for 24 h in U87, U251, LN229, and GBM22 cells. FC: fold change. Shown are means and SD (n = 2–3). ***/**** p < 0.001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32752193), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Bcl-2 Minus C-Terminus Antibody

Application
Recommended Usage

Simple Western

5 µg/mL
Sample: MCF‑7 human breast cancer cell line

Western Blot

0.1-0.5 µg/mL
Sample: KG-1 human acute myelogenous leukemia cell line

Reviewed Applications

Read 3 reviews rated 3.7 using MAB827 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Bcl-2

Bcl-2 is a member of a family of proteins that regulates outer mitochondrial membrane permeability (1, 2). Bcl-2 is an anti-apoptotic member that prevents release of cytochrome c from the mitochondria intermembrane space into the cytosol. Bcl-2 is present on the outer mitochondrial membrane and is also found on other membranes in some cell types. Natural Bcl-2 contains a carboxyl-terminal mitochondria targeting sequence. Recombinant Bcl-2, missing the mitochondrial targeting sequence, maintains its ability to neutralize pro-apoptotic Bcl-2 family members. Neutralization by Bcl-2 appears to be through binding the BH3 region of pro-apoptotic Bcl-2 family members. This activity does not require the mitochondrial targeting sequence.

 

References

  1. Gross, A. et al. (1999) Genes and Develop. 13:1899.
  2. Kroemer, G. (1997) Nature Med. 3:614.

Long Name

B Cell Lymphoma/Leukemia 2

Alternate Names

Bcl2

Entrez Gene IDs

596 (Human); 12043 (Mouse); 24224 (Rat)

Gene Symbol

BCL2

UniProt

Additional Bcl-2 Products

Product Documents for Human Bcl-2 Minus C-Terminus Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Bcl-2 Minus C-Terminus Antibody

For research use only

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