重组Anti-LC3B抗体[EPR18709] - Autophagosome Marker (ab192890)
![Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)](https://www.abcam.cn/ps/products/192/ab192890/Images/ab192890-279980-anti-lc3b-antibody-epr18709-western-blot.jpg "Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18709] to LC3B - Autophagosome Marker
- Suitable for: IP, WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
概述
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产品名称
Anti-LC3B抗体[EPR18709] - Autophagosome Marker
参阅全部 LC3B 一抗 -
描述
兔单克隆抗体[EPR18709] to LC3B - Autophagosome Marker -
宿主
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIP HumanWB MouseRatHuman
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免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: BMDM, U-87 MG, C6 and RAW 264.7 whole cell lysates; Human brain, mouse heart, rat heart, mouse brain and rat brain lysates. IP: HeLa whole cell lysate. ICC/IF: HeLa cells (+/- chloroquine), HAP1 cells (+/- chloroquine) (HAP1-MAP1LC3B knockout cells used as negative cell line).
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
-
形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR18709 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab192890 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| 应用 | Species |
|---|---|
| ICC/IF |
Human
|
| IP |
Human
|
| WB |
Mouse
Rat
Human
|
| 应用 | Ab评论 | 说明 |
|---|---|---|
| IP |
1/50.
|
|
| WB | (8) |
1/2000. Detects a band of approximately 14, 16 kDa (predicted molecular weight: 15 kDa).
|
| ICC/IF | (4) |
Use a concentration of 1 µg/ml.
|
| 说明 |
|---|
|
IP
1/50. |
|
WB
1/2000. Detects a band of approximately 14, 16 kDa (predicted molecular weight: 15 kDa). |
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ICC/IF
Use a concentration of 1 µg/ml. |
靶标
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功能
Probably involved in formation of autophagosomal vacuoles (autophagosomes). -
组织特异性
Most abundant in heart, brain, skeletal muscle and testis. Little expression observed in liver. -
序列相似性
Belongs to the MAP1 LC3 family. -
翻译后修饰
The precursor molecule is cleaved by APG4B/ATG4B to form LC3-I. This is activated by APG7L/ATG7, transferred to ATG3 and conjugated to phospholipid to form LC3-II. -
细胞定位
Cytoplasm > cytoskeleton. Endomembrane system. Cytoplasmic vesicle > autophagosome membrane. LC3-II binds to the autophagic membranes. - Information by UniProt
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数据库链接
- Entrez Gene: 81631 Human
- Entrez Gene: 67443 Mouse
- Entrez Gene: 64862 Rat
- Omim: 609604 Human
- SwissProt: Q9GZQ8 Human
- SwissProt: Q9CQV6 Mouse
- SwissProt: Q62625 Rat
- Unigene: 356061 Human
see all -
别名
- ATG8F antibody
- Autophagy-related protein LC3 B antibody
- Autophagy-related ubiquitin-like modifier LC3 B antibody
see all
图片
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Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: LC3B knockout HAP1 cell lysate (20 µg)
Lane 3: Human brain tissue lysate (20 µg)
Lane 4: U-87 MG cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green).Green -target observed at 14 and 16 kDa. Red - loading control, ab8245, observed at 37 kDa.
This western blot image is a comparison between ab192890 and a competitor's top cited rabbit polyclonal antibody.
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![Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)](https://www.abcam.cn/ps/products/192/ab192890/Images/ab192890-286074-anti-lc3b-antibody-epr18709-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)ab192890 staining LC3B in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells +/- Chloroquine (50μM, 24 hours).
The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab192890 at 1μg/ml and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in pseudocolor red) followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Alexa Fluor® 488 (ab225383) and Alexa Fluor® 647 (ab225382) conjugated versions are available for this clone.
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![Immunoprecipitation - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)](https://www.abcam.cn/ps/products/192/ab192890/Images/ab192890-254798-anti-lc3b-antibody-epr18709-immunoprecipitation.jpg)
Immunoprecipitation - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)LC3B was immunoprecipitated from 1 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab192890 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab192890 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Lane 1: HeLa whole cell lysate 10 µg (Input).
Lane 2: ab192890 IP in HeLa whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR25A] Isotype Control (ab172730) instead of ab192890 in HeLa whole cell cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 30 seconds. -
![Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)](https://www.abcam.cn/ps/products/192/ab192890/Images/ab192890-286072-anti-lc3b-antibody-epr18709-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)ab192890 staining LC3B in HAP1 cells (wildtype and MAP1LC3B knockout) +/- Chloroquine (50μM, 24 hours).
The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab192890 at 1 μg/ml and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in pseudocolor red) followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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![Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)](https://www.abcam.cn/ps/products/192/ab192890/Images/ab192890-266790-anti-lc3b-antibody-epr18709-western-blot.jpg)
Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: LC3B knockout HAP1 cell lysate (20 µg)
Lane 3: Human brain tissue lysate (20 µg)
Lane 4: U-87 MG cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab192890 observed at 14 and 16 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab192890 was shown to specifically react with LC3B in wild-type HAP1 cells. No band was observed when LC3B knockout samples were examined. Wild-type and LC3B knockout samples were subjected to SDS-PAGE. ab192890 and ab8245 (loading control to GAPDH) were diluted 1/2000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging. -
![Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)](https://www.abcam.cn/ps/products/192/ab192890/Images/ab192890-254799-anti-lc3b-antibody-epr18709-western-blot.jpg)
Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)All lanes : Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890) at 1/2000 dilution
Lane 1 : Human brain lysate
Lane 2 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate
Lane 3 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 4 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 5 : Mouse heart lysate
Lane 6 : Rat heart lysate
Lane 7 : Mouse brain lysate
Lane 8 : Rat brain lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 15 kDa
Observed band size: 14,16 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure times: Lane 1-6: 3 minutes; Lane 7 and 8: 30 seconds.
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![Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)](https://www.abcam.cn/ps/products/192/ab192890/Images/ab192890-10-benefits-of-recombinant-antibodies.png)
Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
实验方案
文献 (82)
ab192890 被引用在 82 文献中.
- Cassidy LD et al. Temporal inhibition of autophagy reveals segmental reversal of ageing with increased cancer risk. Nat Commun 11:307 (2020). WB ; Mouse . PubMed: 31949142
- Khorraminejad-Shirazi M et al. AICAR and nicotinamide treatment synergistically augment the proliferation and attenuate senescence-associated changes in mesenchymal stromal cells. Stem Cell Res Ther 11:45 (2020). PubMed: 32014016
- Sasaki N et al. Rapamycin promotes endothelial-mesenchymal transition during stress-induced premature senescence through the activation of autophagy. Cell Commun Signal 18:43 (2020). PubMed: 32164764
- Kong Q et al. Autophagy inhibits TLR4-mediated invasiveness of oral cancer cells via the NF-?B pathway. Oral Dis N/A:N/A (2020). PubMed: 32291890
- Liu H et al. Overexpression of the clock gene Per2 suppresses oral squamous cell carcinoma progression by activating autophagy via the PI3K/AKT/mTOR pathway. J Cancer 11:3655-3666 (2020). PubMed: 32284762
