Anti-VPS35抗体(ab10099)
Key features and details
- Goat polyclonal to VPS35
- Suitable for: IHC-P, WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
- Isotype: IgG
概述
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产品名称
Anti-VPS35抗体
参阅全部 VPS35 一抗 -
描述
山羊多克隆抗体to VPS35 -
宿主
Goat -
经测试应用
适用于: IHC-P, WB, ICC/IFmore details -
种属反应性
与反应: Mouse, Human
预测可用于: Rat, Cow
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免疫原
Synthetic peptide corresponding to Human VPS35 aa 783-796 (C terminal).
Sequence:C-SPESEGPIYEGLIL
(Peptide available asab23181) -
阳性对照
- WB: Human and Mouse Brain lysate. HepG2 and HEK-293 whole cell lysates. IHC-P: Human testis tissue. ICC/IF: U2OS, HeLa, HEK293 cells. IHC-P: Human prostate tissue.
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常规说明
The maintenance of organelles in eukaryotic cells depends on sorting proteins, which ensure the proper delivery of organelle-specific proteins. In S. cerevisiae, VPS35 has a direct role in the retrieval of vacuolar cargo proteins, suggesting that human VPS35 may have an analogous function in the maintenance of lysosomes.
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. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
存储溶液
pH: 7.30
Preservative: 0.02% Sodium azide
Constituents: 99% Tris buffered saline, 0.5% BSA -
Concentration information loading... -
纯度
Immunogen affinity purified -
纯化说明
Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide. -
Primary antibody说明
The maintenance of organelles in eukaryotic cells depends on sorting proteins, which ensure the proper delivery of organelle-specific proteins. In S. cerevisiae, VPS35 has a direct role in the retrieval of vacuolar cargo proteins, suggesting that human VPS35 may have an analogous function in the maintenance of lysosomes. -
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ab10099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 5 - 8 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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| WB | (2) |
Use a concentration of 0.03 - 3 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 91 kDa).
1 hour primary incubation is recommended for this product. |
| ICC/IF | (1) |
Use a concentration of 10 µg/ml.
|
| 说明 |
|---|
|
IHC-P
Use a concentration of 5 - 8 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
|
WB
Use a concentration of 0.03 - 3 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 91 kDa). 1 hour primary incubation is recommended for this product. |
|
ICC/IF
Use a concentration of 10 µg/ml. |
靶标
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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
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数据库链接
- Entrez Gene: 521864 Cow
- Entrez Gene: 100133770 Human
- Entrez Gene: 55737 Human
- Entrez Gene: 65114 Mouse
- Entrez Gene: 25479 Rat
- Omim: 601501 Human
- SwissProt: Q2HJG5 Cow
- SwissProt: Q96QK1 Human
see all -
别名
- DKFZp434E1211 antibody
- DKFZp434P1672 antibody
- FLJ10752 antibody
see all
图片
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Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)Immunocytochemistry/Immunofluorescence analysis of paraformaldehyde fixed U2OS cells, permeabilized with 0.15% Triton. Primary incubation 1hr (10µg/mL) followed by Alexa Fluor 488 secondary antibody (2µg/mL), showing cytoplasmic/vesicle staining. The nuclear stain is DAPI (blue).
Negative control: Unimmunized goat IgG (10µg/mL) followed by Alexa Fluor 488 secondary antibody (2µg/mL). -
Western blot - Anti-VPS35 antibody (ab10099)Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: VPS35 knockout HAP1 cell lysate (20 µg)
Lane 3: NIH3T3 cell lysate (20 µg)
Lane 4: A549 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab10099 observed at 85 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab10099 was shown to specifically react with VPS35 when VPS35 knockout samples were used. Wild-type and VPS35 knockout samples were subjected to SDS-PAGE. ab10099 and ab181602 (loading control to GAPDH) were diluted to 1 µg/ml and 1/1000 respectively and incubated overnight at 4°C. Blots were developed with IRDye® 800CW Donkey anti-Goat IgG (H + L) and Donkey Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216779) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging. -
Western blot - Anti-VPS35 antibody (ab10099)All lanes : Anti-VPS35 antibody (ab10099) at 1 µg/ml
Lane 1 : Human Cerebellum
Lane 2 : Mouse brain
Lysates/proteins at 35 µg per lane.
Predicted band size: 91 kDa -
Western blot - Anti-VPS35 antibody (ab10099)All lanes :
Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate (35µg protein in RIPA buffer).
Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
(35µg protein in RIPA buffer).
Predicted band size: 91 kDaDetected by chemiluminescence.
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Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)Immunofluorescence analysis of paraformaldehyde fixed HEK293 cells, permeabilized with 0.15% Triton. Primary incubation 1hr (10 ug/ml) followed by Alexa Fluor 488 secondary antibody (2 ug/ml), showing cytoplasmic/vesicle staining. The nuclear stain is DAPI (blue).
Negative control: Unimmunized goat IgG (10 ug/ml) followed by Alexa Fluor 488 secondary antibody (2 ug/ml).
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Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)This image is courtesy of an anonymous AbreviewImmunofluorescence analysis of HeLa cells, staining VPS35 with ab10099.
Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 10% serum for 1 hour at 22°C. Samples were incubated with primary antibody (1/150 in 10% Serum in PBS) for 16 hours at 4°C. An AlexaFluor®488-conjugated donkey anti-goat polyclonal IgG (1/300) was used as the secondary antibody. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VPS35 antibody (ab10099)Immunohistochemical analysis of paraffin embedded human prostate tissue labeling VPS35 with ab10099 at 8 ug/ml.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VPS35 antibody (ab10099)Negative Control showing staining of paraffin embedded Human Prostate, with no primary antibody.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (68)
ab10099 被引用在 68 文献中.
- Gopaldass N et al. Retromer oligomerization drives SNX-BAR coat assembly and membrane constriction. EMBO J 42:e112287 (2023). PubMed: 36644906
- Lin G et al. Exploring therapeutic strategies for infantile neuronal axonal dystrophy (INAD/PARK14). Elife 12:N/A (2023). PubMed: 36645408
- Gouveia Roque C et al. CREB3L2-ATF4 heterodimerization defines a transcriptional hub of Alzheimer's disease gene expression linked to neuropathology. Sci Adv 9:eadd2671 (2023). PubMed: 36867706
- Qualls-Histed SJ et al. Lysosomal trafficking of the glucose transporter GLUT1 requires sequential regulation by TXNIP and ubiquitin. iScience 26:106150 (2023). PubMed: 36890792
- Healy MD et al. Structure of the endosomal Commander complex linked to Ritscher-Schinzel syndrome. Cell 186:2219-2237.e29 (2023). PubMed: 37172566