重组Anti-ALKBH5抗体[EPR18958] (ab195377)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18958] to ALKBH5
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-ALKBH5抗体[EPR18958]
参阅全部 ALKBH5 一抗 -
描述
兔单克隆抗体[EPR18958] to ALKBH5 -
宿主
Rabbit -
特异性
This antibody has been tested in IHC-P on Human, Mouse and Rat testis sections. It worked well on Human, but not on the Mouse or Rat sections, under our experimental conditions.
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经测试应用
适用于: WB, IHC-Pmore details -
种属反应性
与反应: Mouse, Rat, Human
预测可用于: Sheep, Goat, Horse, Guinea pig, Cow, Cat, Dog, Pig, Non human primates -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Wild type MEF, HeLa, HepG2 , PC-12, NIH/3T3, Jurkat, Neuro-2a, HEK-293T and F9 whole cell lysates. IHC-P: Human testis and kidney tissues.
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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
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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: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR18958 -
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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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab195377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 |
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WB | (2) |
1/1000. Detects a band of approximately 35-44 kDa (predicted molecular weight: 44 kDa).
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Suitable for human tissues only. |
说明 |
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WB
1/1000. Detects a band of approximately 35-44 kDa (predicted molecular weight: 44 kDa). |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Suitable for human tissues only. |
靶标
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相关性
Dioxygenase that demethylates RNA by oxidative demethylation: specifically demethylates N(6)-methyladenosine (m6A) RNA, the most prevalent internal modification of messenger RNA (mRNA) in higher eukaryotes. Requires molecular oxygen, alpha-ketoglutarate and iron. Demethylation of m6A mRNA affects mRNA processing and export and is required for spermatogenesis. -
细胞定位
Nucleus speckle -
数据库链接
- Entrez Gene: 533303 Cow
- Entrez Gene: 54890 Human
- Entrez Gene: 268420 Mouse
- Entrez Gene: 303193 Rat
- SwissProt: E1BH29 Cow
- SwissProt: Q6P6C2 Human
- SwissProt: Q3TSG4 Mouse
- SwissProt: D3ZKD3 Rat
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别名
- ABH5 antibody
- AlkB, alkylation repair homolog 5 (E. coli) antibody
- AlkB, alkylation repair homolog 5 antibody
see all
图片
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All lanes : Anti-ALKBH5 antibody [EPR18958] (ab195377) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : ALKBH5 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 44 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab195377 observed at 48 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab195377 was shown to react with ALKBH5 in wild-type HEK-293T cells in western blot with loss of signal observed in ALKBH5 knockout cell line ab266762 (ALKBH5 knockout cell lysate ab257349). Wild-type and ALKBH5 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab195377 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemical analysis of paraffin-embedded human testis tissue labeling ALKBH5 with ab195377 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in human testis (PMID: 21264265; PMID: 27590511; PMID: 17979886) is observed. Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
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Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling ALKBH5 with ab195377 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in human kidney (PMID: 21264265) is observed. Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
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Lanes 1-2 : Anti-ALKBH5 antibody [EPR18958] (ab195377) at 1/1000 dilution
Lanes 3-5 : Anti-ALKBH5 antibody [EPR18958] (ab195377) at 1/2000 dilution
Lane 1 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 2 : NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate
Lane 3 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 4 : Neuro-2a (mouse neuroblastoma cell line) whole cell lysate
Lane 5 : F9 (mouse embryonic testicular cancer cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 44 kDa
Observed band size: 35-44 kDa why is the actual band size different from the predicted?Exposure time : Lanes 1 and 2: 15 seconds; Lanes 3 and 4: 26 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
The molecular mass observed is consistent with the literature (PMID 21264265).
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All lanes : Anti-ALKBH5 antibody [EPR18958] (ab195377) at 1/5000 dilution
Lane 1 : Wild type MEF whole cell lysate
Lane 2 : ALKBH5 knockout MEF whole cell lysate
Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 44 kDa
Observed band size: 35-44 kDa why is the actual band size different from the predicted?
Exposure time: 70 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The Wt and ALKBH5 KO cell lysates were kindly provided by an anonymous collaborator.
The molecular mass observed is consistent with the literature (PMID 21264265).
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (52)
ab195377 被引用在 52 文献中.
- Zhang H et al. A novel molecular classification method for osteosarcoma based on tumor cell differentiation trajectories. Bone Res 11:1 (2023). IHC-P ; Human . PubMed: 36588108
- Wang W et al. ALKBH5 prevents hepatocellular carcinoma progression by post-transcriptional inhibition of PAQR4 in an m6A dependent manner. Exp Hematol Oncol 12:1 (2023). IHC-P ; Human . PubMed: 36609413
- Ge J et al. RNA demethylase ALKBH5 suppresses tumorigenesis via inhibiting proliferation and invasion and promoting CD8+ T cell infiltration in colorectal cancer. Transl Oncol 34:101683 (2023). WB ; Human . PubMed: 37224767
- Li Y et al. Post-transcriptional modification of m6A methylase METTL3 regulates ERK-induced androgen-deprived treatment resistance prostate cancer. Cell Death Dis 14:289 (2023). IHC-P ; Human . PubMed: 37095108
- Chen J et al. Inhibition of ALKBH5 attenuates I/R-induced renal injury in male mice by promoting Ccl28 m6A modification and increasing Treg recruitment. Nat Commun 14:1161 (2023). WB, IHC-P ; Mouse . PubMed: 36859428