Anti-DCAMKL1抗体(ab31704)
Key features and details
- Rabbit polyclonal to DCAMKL1
- Suitable for: ICC/IF, WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
选择批间可重复性更高的重组抗体
- 研究可靠 —— 各批次间结果一致且可重复
- 长期批量供应 —— 采用重组技术，可实现快速生产
- 首次实验即可成功 —— 经过大量验证确认了特异性
- 符合伦理标准 —— 产品不含动物成分
概述
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产品名称
Anti-DCAMKL1抗体
参阅全部 DCAMKL1 一抗 -
描述
兔多克隆抗体to DCAMKL1 -
宿主
Rabbit -
特异性
This antibody recognizes 2 different human isoforms (AL: 82.2 KDa and BL: 47.6 KDa). From May 2024, QC testing of replenishment batches of this polyclonal changed. All tested and expected application and reactive species combinations are still covered by our Abcam product promise. However, we no longer test all applications. For more information on a specific batch, please contact our Scientific Support who will be happy to help. You may also be interested in our alternative recombinant antibody, ab109029.
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经测试应用
适用于: ICC/IF, WB, IPmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HAP1 and NIH-3T3 cell lysates. Human, mouse, and rat brain tissue lysates;
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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
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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 or -80°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
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
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Compatible Secondaries
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab31704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 |
|---|---|---|
| ICC/IF | (1) |
Use a concentration of 5 µg/ml.
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| WB | (4) |
Use a concentration of 0.5 µg/ml. Detects a band of approximately 47, 82 kDa (predicted molecular weight: 47, 82 kDa).
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| IP |
Use at an assay dependent concentration.
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| 说明 |
|---|
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ICC/IF
Use a concentration of 5 µg/ml. |
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WB
Use a concentration of 0.5 µg/ml. Detects a band of approximately 47, 82 kDa (predicted molecular weight: 47, 82 kDa). |
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IP
Use at an assay dependent concentration. |
靶标
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功能
Probable kinase that may be involved in a calcium-signaling pathway controlling neuronal migration in the developing brain. May also participate in functions of the mature nervous system. -
组织特异性
In fetal tissues, highly expressed in brain, detectable in lung and liver, but not in kidney. In adult tissues, expressed ubiquitously in the brain, detectable in the heart, liver, spleen, thymus, prostate, testis, ovary, small intestine and colon. The type A isoforms seem to be expressed predominantly in fetal brain whereas type B isoforms are expressed abundantly in both fetal and adult brain. -
序列相似性
Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CaMK subfamily.
Contains 2 doublecortin domains.
Contains 1 protein kinase domain. - Information by UniProt
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数据库链接
- Entrez Gene: 9201 Human
- Entrez Gene: 13175 Mouse
- Entrez Gene: 83825 Rat
- Omim: 604742 Human
- SwissProt: O15075 Human
- SwissProt: Q9JLM8 Mouse
- SwissProt: O08875 Rat
- Unigene: 507755 Human
see all -
别名
- Calcium/calmodulin-dependent protein kinase type I-like CPG16 antibody
- CL1 antibody
- CLICK1 antibody
see all
图片
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Western blot - Anti-DCAMKL1 antibody (ab31704)This image is courtesy of an anonymous AbreviewAll lanes : Anti-DCAMKL1 antibody (ab31704) at 1 µg/ml
Lane 1 : HCT116 WT
Lane 2 : MiaPaCa2
Lane 3 : SW480
Lane 4 : SW620
Lane 5 : HT-29
Lane 6 : H4
Lane 7 : mouse brain lysate
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : IRDye® 800CW Goat Anti-Rabbit at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 47, 82 kDa
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Immunocytochemistry/ Immunofluorescence - Anti-DCAMKL1 antibody (ab31704)ab31704 staining DCAMKL1 in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween 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 ab31704 at 5 µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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Western blot - Anti-DCAMKL1 antibody (ab31704)Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: DCAMKL1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: NIH3T3 whole cell lysate (20 µg)
Lane 4: Human brain whole tissue lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab31704 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab31704 was shown to specifically react with DCAMKL1 in wild-type HAP1 cells along with additional cross reactive bands. No bands were observed when DCAMKL1 knockout cells were examined. Wild-type and DCAMKL1 knockout samples were subjected to SDS-PAGE. ab31704 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 µg/ml and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
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Immunoprecipitation - Anti-DCAMKL1 antibody (ab31704)DCAMKL1 was immunoprecipitated using 0.5mg Mouse Brain whole tissue lysate, 5µg of Rabbit polyclonal to DCAMKL1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab31704.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 82kDa: DCAMKL1; non specific - 52 and 27kDa: We are unsure as to the identity of this extra band. -
Western blot - Anti-DCAMKL1 antibody (ab31704)Anti-DCAMKL1 antibody (ab31704) at 1 µg/ml + Mouse Brain Whole Tissue Lysate at 20 µg
Secondary
IR Dye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution
Performed under reducing conditions.
Predicted band size: 47, 82 kDa
Observed band size: 47,82 kDa why is the actual band size different from the predicted?
The 82 kDa and 47 kDa bands correspond to the AL and BL isoforms respectively. This antibody should not detect the AS and BS isoforms of DCAMLK1. -
Immunocytochemistry/ Immunofluorescence - Anti-DCAMKL1 antibody (ab31704)ICC/IF image of ab31704 stained PC12 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab31704, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue). -
Western blot - Anti-DCAMKL1 antibody (ab31704)Anti-DCAMKL1 antibody (ab31704) at 1 µg/ml + Brain (Rat) Tissue Lysate - normal tissue at 10 µg
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 47, 82 kDa
Observed band size: 47,82 kDa why is the actual band size different from the predicted? -
Western blot - Anti-DCAMKL1 antibody (ab31704)Anti-DCAMKL1 antibody (ab31704) 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
Performed under reducing conditions.
Predicted band size: 47, 82 kDa
Observed band size: 47,82 kDa why is the actual band size different from the predicted?
Additional bands at: 30 kDa, 52 kDa (possible post-translational modification), 54 kDa (possible post-translational modification). We are unsure as to the identity of these extra bands.
Exposure time: 5 minutes
The 82 kDa and 47 kDa bands correspond to the AL and BL isoforms respectively. DCAMLK1 has a number of potential phosphorylation sites which may explain the higher migrating bands at 52 and 54 kDa.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (129)
ab31704 被引用在 129 文献中.
- Huang Z et al. Macrophage DCLK1 promotes atherosclerosis via binding to IKKβ and inducing inflammatory responses. EMBO Mol Med 15:e17198 (2023). PubMed: 36896602
- Lucas B et al. Embryonic keratin19+ progenitors generate multiple functionally distinct progeny to maintain epithelial diversity in the adult thymus medulla. Nat Commun 14:2066 (2023). PubMed: 37045811
- Liu H et al. Targeting DCLK1 attenuates tumor stemness and evokes antitumor immunity in triple-negative breast cancer by inhibiting IL-6/STAT3 signaling. Breast Cancer Res 25:43 (2023). PubMed: 37069669
- Cui W et al. Diet-mediated constitutive induction of novel IL-4+ ILC2 cells maintains intestinal homeostasis in mice. J Exp Med 220:N/A (2023). PubMed: 37163450
- Klein F et al. Combined multidimensional single-cell protein and RNA profiling dissects the cellular and functional heterogeneity of thymic epithelial cells. Nat Commun 14:4071 (2023). PubMed: 37429879