Human/Mouse/Rat Bcl-2 Antibody

Catalog # Availability Size / Price Qty
MAB8272
MAB8272-SP
Detection of Human, Mouse, and Rat Bcl‑2 by Western Blot.
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Product Details
Citations (12)
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Human/Mouse/Rat Bcl-2 Antibody Summary

Species Reactivity
Human, Mouse, Rat
Specificity
Detects human Bcl-2 in direct ELISAs and Western blots. In direct ELISAs, approximately 25‑40% cross‑reactivity with recombinant mouse (rm) Bcl‑2 is observed and no cross-reactivity with recombinant human BCL2L12 is observed. In Western blots, 100% cross-reactivity with rmBcl-2 is observed.
Source
Monoclonal Mouse IgG2B Clone # 625509
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
E. coli-derived recombinant human Bcl-2
Ala2-Asp211
Accession # P10415
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
See below
Immunohistochemistry
8-25 µg/mL
See below
Knockout Validated
Bcl‑2 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in Bcl‑2 knockout HeLa cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human, Mouse, and Rat Bcl-2 antibody by Western Blot. View Larger

Detection of Human, Mouse, and Rat Bcl‑2 by Western Blot. Western blot shows lysates of THP-1 human acute monocytic leukemia cell line, KG-1 human acute myelogenous leukemia cell line, CTLL-2 mouse cytotoxic T cell line, and NRK rat normal kidney cell line. PVDF Membrane was probed with 0.1 µg/mL of Mouse Anti-Human/Mouse/Rat Bcl-2 Monoclonal Antibody (Catalog # MAB8272) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Bcl-2 at approximately 24 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry Bcl-2 antibody in Human Lymphoma by Immunohistochemistry (IHC-P). View Larger

Bcl‑2 in Human Lymphoma. Bcl-2 was detected in immersion fixed paraffin-embedded sections of human lymphoma using Mouse Anti-Human/Mouse/Rat Bcl-2 Monoclonal Antibody (Catalog # MAB8272) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Knockout Validated Western Blot Shows Human Bcl-2 Antibody Specificity by Using Knockout Cell Line. View Larger

Western Blot Shows Human Bcl‑2 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Bcl-2 knockout HeLa cell line (KO). PVDF membrane was probed with 0.1 µg/mL of Mouse Anti-Human/Mouse/Rat Bcl-2 Monoclonal Antibody (Catalog # MAB8272) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Bcl-2 at approximately 24 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Western Blot Detection of Human Bcl-2 by Western Blot View Larger

Detection of Human Bcl-2 by Western Blot Cholesterol reversed metformin-mediated anticancer potential.Cells were treated with metformin (2 mM) alone and in combination with metformin (2 mM) plus cholesterol (10 μg/ml). (A) Cell viability (MTT) assay was performed to check the reverse effect of cholesterol on metformin- inhibited cell viability. Bars are representing the relative cell viability after 24 hr of treatment. Values represent mean ± SEM of triplicate measurements, **p <.001 vs. control and *p <.001 vs. metformin. (B) Scratch assay was performed. MDA-MB-231 cell monolayers were scratched and incubated with metformin (2 mM) alone and in combination with metformin (2 mM) plus cholesterol (10 μg/ml). The cell monolayers were photographed, and represented photos at 0 hr and 24 hr for control, metformin and metformin plus cholesterol cells were shown here. (C) The unfilled gap areas between two ends of the scratch were measured and subsequently plotted. Values represent mean ± SEM of triplicate measurements, **p <.01 vs. control at 0 hr and **p <.01 vs. metformin at 24 hr. (D) Colony formation assay was performed. After 24 hr of cell seeding, cells were treated with metformin (1mM) alone and in combination with metformin (1mM) plus cholesterol (5 μg/ml). After 5 days, formed colonies were stained with crystal violet. Photos of the wells were taken by a camera. (E) Soft agar assay of MDA-MB-231 cells in presence and absence of metformin (1mM) alone and in combination with metformin (1mM) plus cholesterol (5 μg/ml). Day 9 pictures were shown in the figure. Arrows depict the spheres. (F, G and H) RT-PCR analysis was performed using total RNA isolated from metformin, metformin (1mM) plus cholesterol treated and untreated cells, and gene specific primers. Cholesterol treatment reverses the metformin inhibited genes expressions (Bcl-xL [F], Zeb1 [G] and BMI1 [H]) (compare lane 2 vs. lane 3). (I and J) Western blot analysis was performed using total proteins from metformin, metformin (1mM) plus cholesterol treated and untreated cells, and specific antibodies. Cholesterol treatment reverses the metformin inhibited genes expression (BCL2 [I] and vimentin [J]) (compare lane 2 vs. lane 3). (K) Gelatin zymography was performed using equal amount of conditioned medium of metformin, metformin (2 mM) plus cholesterol (10 μg/ml) treated and untreated cells. Metformin-inhibited MMP activity was increased by treatment of cholesterol (compare lane 2 vs. lane 3). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30625181), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.5 mg/mL.
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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
Entrez Gene IDs
596 (Human); 12043 (Mouse); 24224 (Rat)
Alternate Names
apoptosis regulator Bcl-2; B-cell CLL/lymphoma 2; Bcl2; Bcl-2

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Citations for Human/Mouse/Rat Bcl-2 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

12 Citations: Showing 1 - 10
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  1. DL-3-n-butylphthalide protects endothelial cells against advanced glycation end product-induced injury by attenuating oxidative stress and inflammation responses
    Authors: Chang-Yun Liu, Zhen-Hua Zhao, Zhi-Ting Chen, Chun-Hui Che, Zhang-Yu Zou, Xiao-Min Wu et al.
    Experimental and Therapeutic Medicine
  2. Metformin induces pyroptosis in leptin receptor-defective hepatocytes via overactivation of the AMPK axis
    Authors: B Liu, J Xu, L Lu, L Gao, S Zhu, Y Sui, T Cao, T Yang
    Cell Death & Disease, 2023-02-03;14(2):82.
    Species: Rat
    Sample Types: Cell Lysates
    Applications: Western Blot
  3. Human Umbilical Cord Mesenchymal Stem Cells Attenuate Severe Burn-Induced Multiple Organ Injury via Potentiating IGF-1 and BCL-2/BAX Pathway
    Authors: H Wang, T Ba, Q Wang, L Yang, C Li, X Hao, Y Yin, L Liu
    Stem Cells International, 2022-12-23;2022(0):5474289.
    Species: Rat
    Sample Types: Tissue Lysates
    Applications: Western Blot
  4. Cyclo(Val-Pro) and Cyclo(Leu-Hydroxy-Pro) from Pseudomonas sp. (ABS-36) alleviates acute and chronic renal injury under in vitro and in vivo models (Ischemic reperfusion and unilateral ureter obstruction)
    Authors: K Hira, P Sharma, A Mahale, O Prakash Ku, A Sajeli Beg
    International immunopharmacology, 2021-12-29;103(0):108494.
    Species: Rat
    Sample Types: Cell Lysates
    Applications: Western Blot
  5. Curcumin alleviates arsenic-induced toxicity in PC12�cells via modulating autophagy/apoptosis
    Authors: MS Rahaman, S Banik, M Akter, MM Rahman, MT Sikder, T Hosokawa, T Saito, M Kurasaki
    Ecotoxicol. Environ. Saf., 2020-05-25;200(0):110756.
    Species: Rat
    Sample Types: Whole Cell
    Applications: Cell Culture
  6. Metformin exhibited anticancer activity by lowering cellular cholesterol content in breast cancer cells
    Authors: A Sharma, S Bandyopadh, K Chowdhury, T Sharma, R Maheshwari, A Das, G Chakrabart, V Kumar, CC Mandal
    PLoS ONE, 2019-01-09;14(1):e0209435.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  7. Dexmedetomidine protects the developing rat brain against the neurotoxicity wrought by sevoflurane: role of autophagy and Drp1-Bax signaling
    Authors: Y Shan, S Sun, F Yang, N Shang, H Liu
    Drug Des Devel Ther, 2018-10-30;12(0):3617-3624.
    Species: Rat
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  8. FK506 Attenuates the MRP1-Mediated Chemoresistant Phenotype in Glioblastoma Stem-Like Cells
    Authors: Á Torres, V Arriagada, JI Erices, MLÁ Toro, JD Rocha, I Niechi, C Carrasco, C Oyarzún, C Quezada
    Int J Mol Sci, 2018-09-11;19(9):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  9. Exosomes derived from human umbilical cord mesenchymal stem cells improve myocardial repair via upregulation of Smad7
    Authors: XL Wang, YY Zhao, L Sun, Y Shi, ZQ Li, XD Zhao, CG Xu, HG Ji, M Wang, WR Xu, W Zhu
    Int. J. Mol. Med., 2018-02-16;0(0):.
    Species: Human, Rat
    Sample Types: Cell Lysates, Tissue Homogenates
    Applications: Western Blot
  10. Hepatoprotective effect of calculus bovis sativus on nonalcoholic fatty liver disease in mice by inhibiting oxidative stress and apoptosis of hepatocytes
    Authors: W He, Y Xu, C Zhang, J Lu, J Li, D Xiang, J Yang, M Chang, D Liu
    Drug Des Devel Ther, 2017-12-05;11(0):3449-3460.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Western Blot

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