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免疫原 :E.coli-derived human Beclin 1 recombinant protein (Position: M1-S354). Human Beclin 1 shares 97% amino acid (aa) sequence identity with both mouse and rat Beclin 1.
亚型 :Rabbit IgG
形态 :Lyophilized
保存条件 :At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time. Avoid repeated freezing and thawing.
克隆性 :单克隆
适应物种 :Human,Mouse,Rat
抗原来源 :Rabbit
库存 :现货
供应商 :博士德生物
应用范围 :WB,IHC-P,FCM
浓度 :Concentration: 0.1-0.5μg/ml; Tested Species: Human
规格 :100ul/200ul
产品概况
货号 | PB0014 |
---|---|
产品名称 | ANTI-BECN1 Antibody |
基因名 | BECN1 |
抗体来源 | Rabbit |
克隆 | Polyclonal |
抗体亚型 | Rabbit IgG |
免疫原 | E.coli-derived human Beclin 1 recombinant protein (Position: M1-S354). Human Beclin 1 shares 97% amino acid (aa) sequence identity with both mouse and rat Beclin 1. |
内容 | 500ug/ml antibody with PBS ,0.02% NaN3 , 1mg BSA |
纯化方式 | Immunogen affinity purified. |
浓度 | Concentration: 0.1-0.5μg/ml; Tested Species: Human |
产品形态 | Lyophilized |
保存条件 | At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time. Avoid repeated freezing and thawing. |
背景资料 | Beclin-1, also known as also known as ATG6 or VPS30 is a protein that in humans is encoded by the BECN1 gene. Beclin-1 and its binding partner class III phosphoinositide 3-kinase (PI3K), also named Vps34, are required for the initiation of the formation of the autophagosome in autophagy. This gene participates in the regulation of autophagy and has an important role in development, tumorigenesis, and neurodegeneration. Schizophrenia is associated with low levels of Beclin-1 in the hippocampus of the affected which causes diminished autophagywhich in turn results in increased neuronal cell death. It has been found that beclin-1 can promote autophagy in autophagy-defective yeast with a targeted disruption of apg6/vps30, and in human MCF7 breast carcinoma cells. |
研究类别 | 1. Liang, X. H., Kleeman, L. K., Jiang, H. H., Gordon, G., Goldman, J. E., Berry, G., Herman, B., Levine, B.Protection against fatal Sindbis virus encephalitis by beclin, a novel Bcl-2-interacting protein. J. Virol. 72: 8586-8596, 1998.2. Merenlender-Wagner A, Malishkevich A, Shemer Z, Udawela M, Gibbons A, Scarr E, Dean B, Levine J, Agam G, Gozes I (December 2013). "Autophagy has a key role in the pathophysiology of schizophrenia". Mol. Psychiatry. |
Uniprot ID | BECN1: Q14457 |
推荐配套的二抗和检测试剂 | Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P). *Blocking peptide 可以联系我们购买。 |
产品应用细节
为了提供最优质的抗体,博士德对每一批抗体都用没有转染过的细胞系和体细胞组织检测,以保证博士德出品的抗体有足够的亲和性足以和对应蛋白天然的表达含量起反应。
应用 | 稀释度* |
---|---|
Western blot: | 1:500-2000 |
Immunohistochemistry in paraffin section: | 1:50-400 |
Flow Cytometry: | 1-3μg/1x106 cells |
(Boiling the paraffin sections in 10mM citrate buffer,pH6.0,or PH8.0 EDTA repair liquid for 20 mins is required for the staining of formalin/paraffin sections.) Optimal working dilutions must be determined by end user. |
*最优稀释度需要用户自己调试,此处数据仅供参考。
**博士德提供高敏感的二抗和检测试剂盒。详情见相关产品推荐。
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[list_product_images]Figure 1. IHC analysis of Beclin-1 using anti-Beclin-1 antibody (PB9076).Beclin-1 was detected in paraffin-embedded section of Mouse Intestine Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Beclin-1 Antibody (PB9076) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen. |Figure 2. IHC analysis of Beclin-1 using anti-Beclin-1 antibody (PB9076).Beclin-1 was detected in paraffin-embedded section of Mouse Spleen Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Beclin-1 Antibody (PB9076) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.|Figure 3. IHC analysis of Beclin-1 using anti-Beclin-1 antibody (PB9076).Beclin-1 was detected in paraffin-embedded section of Rat Intestine Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Beclin-1 Antibody (PB9076) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.|Figure 4. IHC analysis of Beclin-1 using anti-Beclin-1 antibody (PB9076).Beclin-1 was detected in paraffin-embedded section of Rat Spleen Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Beclin-1 Antibody (PB9076) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.|Figure 5. Western blot analysis of Beclin-1 using anti-Beclin-1 antibody (PB9076). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: Colo320 Whole Cell LysateLane 2: HepG2 Whole Cell LysateLane 3: PANC Whole Cell LysateLane 4: A431 Whole Cell LysateLane 5: Smmc Whole Cell LysateLane 6: Jurkat Whole Cell LysateLane 7: SW620 Whole Cell LysateLane 8: U87 Whole Cell LysateAfter Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Beclin-1 antigen affinity purified polyclonal antibody (Catalog # PB9076) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Beclin-1 at approximately 60KD. The expected band size for Beclin-1 is at 52KD.|Figure 6. Flow Cytometry analysis of HepG2 cells using anti- Beclin-1 antibody (PB9076).
Overlay histogram showing HepG2 cells stained with PB9076 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti- Beclin-1 Antibody (PB9076,1μg/1x106 cells) for 30 min at 20°C. DyLight 488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.[/list_product_images]
Overlay histogram showing HepG2 cells stained with PB9076 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti- Beclin-1 Antibody (PB9076,1μg/1x106 cells) for 30 min at 20°C. DyLight 488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.[/list_product_images]
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