Anti-VPS35抗体[2D3] (ab57632)
Key features and details
- Mouse monoclonal [2D3] to VPS35
- Suitable for: WB, ICC/IF, Flow Cyt
- Knockout validated
- Reacts with: Human, Recombinant fragment
- Isotype: IgG2a
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
-
产品名称
Anti-VPS35抗体[2D3]
参阅全部 VPS35 一抗 -
描述
小鼠单克隆抗体[2D3] to VPS35 -
宿主
Mouse -
经测试应用
适用于: WB, ICC/IF, Flow Cytmore details -
种属反应性
与反应: Human, Recombinant fragment -
免疫原
Recombinant fragment: MECLKKALKI ANQCMDPSLQ VQLFIEILNR YIYFYEKEND AVTIQVLNQL IQKIREDLPN LESSEETEQI NKHFHNTLEH LRLRRESPES EGPIYEGLIL , corresponding to amino acids 697-797 of Human VPS35
-
阳性对照
- Flow Cyt: HEK293 cells. WB: Wild-type HAP1, NIH/3T3, A549 cell lysates, tagged recombinant protein. ICC/IF: Wild-type HAP1 cells.
-
常规说明
This product was changed from ascites to tissue culture supernatant on 05 Feb 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
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. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
存储溶液
pH: 7.4 -
Concentration information loading...
-
纯度
Tissue culture supernatant -
纯化说明
Purified from TCS. -
克隆
单克隆 -
克隆编号
2D3 -
同种型
IgG2a -
轻链类型
kappa -
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ab57632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 |
---|---|---|
WB |
Use at an assay dependent concentration.
This antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein. |
|
ICC/IF |
Use a concentration of 0.2 µg/ml.
|
|
Flow Cyt |
Use at an assay dependent concentration.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
说明 |
---|
WB
Use at an assay dependent concentration. This antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein. |
ICC/IF
Use a concentration of 0.2 µg/ml. |
Flow Cyt
Use at an assay dependent concentration. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
靶标
-
功能
Essential component of the retromer complex, a complex required to retrieve lysosomal enzyme receptors (IGF2R and M6PR) from endosomes to the trans-Golgi network. Also required to regulate transcytosis of the polymeric immunoglobulin receptor (pIgR-pIgA). -
组织特异性
Ubiquitous. Highly expressed in heart, brain, placenta, skeletal muscle, spleen, thymus, testis, ovary, small intestine, kidney and colon. -
序列相似性
Belongs to the VPS35 family. -
细胞定位
Cytoplasm. Membrane. - Information by UniProt
-
数据库链接
- Entrez Gene: 100133770 Human
- Entrez Gene: 55737 Human
- Omim: 601501 Human
- SwissProt: Q96QK1 Human
- Unigene: 454528 Human
-
别名
- DKFZp434E1211 antibody
- DKFZp434P1672 antibody
- FLJ10752 antibody
see all
图片
-
Ab57632 staining VPS35 in wild-type HAP1 cells (top panel) and VPS35 knockout HAP1 cells (bottom panel). The cells were fixed with 4% PFA (10 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 with ab57632 at 0.2 µg/mL and ab6046 at 1 µg/mL overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Mouse IgG (Alexa Fluor® 488) (ab150117) (shown in pseudo colour green) and goat secondary antibody to Rabbit IgG (Alexa Fluor® 594) (ab150084) (shown in pseudo colour red) both at 1/1000. Nuclear DNA was labelled with DAPI (shown in pseudo colour blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
-
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: VPS35 knockout HAP1 cell lysate (20 µg)
Lane 3: NIH/3T3 cell lysate (20 µg)
Lane 4: A549 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab57632 observed at 91 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab57632 was shown to specifically react with VPS35 in wild-type HAP1 cells. No band was observed when VPS35 knockout samples were examined. Wild-type and VPS35 knockout samples were subjected to SDS-PAGE. ab5732 and ab181602 (loading control to GAPDH) were diluted 1µg/mL and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.This image was generated using a version of the antibody produced in ascites.
-
Western blot against tagged recombinant protein immunogen using ab57632 VPS35 antibody at 1 μg/ml. Predicted band size of immunogen is 37 kDa
This image was generated using a version of the antibody produced in ascites.
-
Overlay histogram showing HEK293 cells stained with ab57632 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab57632, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
This image was generated using a version of the antibody produced in ascites.
数据表及文件
-
Datasheet download
文献 (18)
ab57632 被引用在 18 文献中.
- Qureshi YH et al. The neuronal retromer can regulate both neuronal and microglial phenotypes of Alzheimer's disease. Cell Rep 38:110262 (2022). PubMed: 35045281
- Wilhelmi I et al. The ARFRP1-dependent Golgi scaffolding protein GOPC is required for insulin secretion from pancreatic ß-cells. Mol Metab 45:101151 (2021). PubMed: 33359402
- Sargent D et al. Neuronal VPS35 deletion induces spinal cord motor neuron degeneration and early post-natal lethality. Brain Commun 3:fcab208 (2021). PubMed: 34704029
- Xie J et al. TBC1D5-Catalyzed Cycling of Rab7 Is Required for Retromer-Mediated Human Papillomavirus Trafficking during Virus Entry. Cell Rep 31:107750 (2020). PubMed: 32521275
- Simoes S et al. Tau and other proteins found in Alzheimer's disease spinal fluid are linked to retromer-mediated endosomal traffic in mice and humans. Sci Transl Med 12:N/A (2020). PubMed: 33239387