Anti-LC3B抗体(ab63817)
Key features and details
- Rabbit polyclonal to LC3B
- Suitable for: IHC-P, WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
概述
-
产品名称
Anti-LC3B抗体
参阅全部 LC3B 一抗 -
描述
兔多克隆抗体to LC3B -
宿主
Rabbit -
经测试应用
适用于: IHC-P, WB, ICC/IFmore details -
种属反应性
与反应: Mouse, Rat, Human
预测可用于: Chicken, Cow, Macaque monkey, Zebra finch -
免疫原
-
阳性对照
- This antibody gave a positive signal in Human, Mouse and Rat Brain Tissue Lysate. ICC/IF: HeLa cells (+/- chloroquine), HAP1 cells (+/-chloroquine) (HAP1-MAP1LC3B knockout cells used as negative cell line).
-
常规说明
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
-
纯度
Immunogen affinity purified -
克隆
多克隆 -
同种型
IgG -
研究领域
相关产品
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab63817于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
---|---|---|
IHC-P |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
|
WB |
Use a concentration of 1 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
|
|
ICC/IF | (1) |
Use a concentration of 1 µg/ml.
|
说明 |
---|
IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa). |
ICC/IF
Use a concentration of 1 µg/ml. |
靶标
-
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
-
数据库链接
- Entrez Gene: 427559 Chicken
- Entrez Gene: 408001 Cow
- Entrez Gene: 81631 Human
- Entrez Gene: 67443 Mouse
- Entrez Gene: 64862 Rat
- Omim: 609604 Human
- SwissProt: O41515 Cow
- SwissProt: Q9GZQ8 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
-
All lanes : Anti-LC3B antibody (ab63817) at 1 µg/ml
Lane 1 : Wild-type HepG2 untreated control cell lysate
Lane 2 : Wild-type HepG2 Treated Chloroquine (50 uM, 16 h) cell lysate
Lane 3 : MAP1LC3B knockout HepG2 untreated control cell lysate
Lane 4 : MAP1LC3B knockout HepG2 Treated Chloroquine (50 uM, 16 h) cell lysate
Lane 5 : U-87 MG cell lysate
Lane 6 : PC-12 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 14,16 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-LC3B antibody staining at 1 ug/ml, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab63817 was shown to bind specifically to LC3B. A band was observed at 16/14 kDa (yellow arrows) in treated wild-type HepG2 cell lysates with no signal observed at this size in MAP1LC3B knockout cell line ab277828 (knockout cell lysate ab283796). To generate this image, wild-type and MAP1LC3B knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
-
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: U87MG cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab63817 observed at 16 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab63817 was shown to recognize LC3B in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when LC3B knockout samples were examined. Wild-type and LC3B knockout samples were subjected to SDS-PAGE. ab63817 and ab8245 (loading control to GAPDH) were diluted 1 µg/mL 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. -
ab63817 staining LC3B in HeLa 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 ab63817 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 labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
-
ab63817 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 ab63817 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 labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
-
IHC image of LC3B staining in human hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab63817, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
-
Anti-LC3B antibody (ab63817) at 1 µg/ml + Human brain tissue lysate - total protein (ab29466) at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 15 kDa
Additional bands at: 32 kDa, 54 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 2 minutes -
All lanes : Anti-LC3B antibody (ab63817) at 1 µg/ml
Lane 1 : Brain (Mouse) Tissue Lysate
Lane 2 : Brain (Rat) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 15 kDa
Additional bands at: 30 kDa, 65 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 3 minutes -
Immunocytochemical analysis of HeLa cells, labeling LC3B with ab63817 (1/200 with PBS for 16 hours at 22°C). Cells were methanol fixed, immunofluorescently labeled, adn counterstained with DAPI.
实验方案
数据表及文件
-
SDS download
-
Datasheet download
文献 (60)
ab63817 被引用在 60 文献中.
- Kong M et al. The dynamic changes in autophagy activity and its role in lung injury after deep hypothermic circulatory arrest. J Cell Mol Med 26:1113-1127 (2022). PubMed: 35014165
- Duan M et al. Mitochondrial apolipoprotein A-I binding protein alleviates atherosclerosis by regulating mitophagy and macrophage polarization. Cell Commun Signal 20:60 (2022). PubMed: 35525979
- Li L et al. Luteolin alleviates inflammation and autophagy of hippocampus induced by cerebral ischemia/reperfusion by activating PPAR gamma in rats. BMC Complement Med Ther 22:176 (2022). PubMed: 35778706
- Liu B et al. Locally generated C3 regulates the clearance of Toxoplasma gondii by IFN-γ-primed macrophage through regulation of xenophagy. Front Microbiol 13:944006 (2022). PubMed: 35992649
- Huang X et al. PRRX1/FOXM1 reduces gemcitabin-induced cytotoxicity by regulating autophagy in bladder cancer. Transl Androl Urol 11:1116-1129 (2022). PubMed: 36092834