Anti-VPS35抗体
参阅全部 VPS35 一抗
山羊多克隆抗体to VPS35
Goat
适用于: IHC-P, WB, ICC/IFmore details
与反应: Mouse, Human
预测可用于: Rat, Cow
Synthetic peptide corresponding to Human VPS35 aa 783-796 (C terminal).
Sequence:
C-SPESEGPIYEGLIL
(Peptide available as ab23181)
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.
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.
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Liquid
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
pH: 7.30
Preservative: 0.02% Sodium azide
Constituents: 99% Tris buffered saline, 0.5% BSA
浓度
100 µg 浓度为 0.5 mg/ml
Immunogen affinity purified
Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
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.
多克隆
IgG
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. | |
| 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. |
Entrez Gene: 100133770 Human
Entrez Gene: 55737 Human
Entrez Gene: 65114 Mouse
Omim: 601501 Human
SwissProt: Q96QK1 Human
SwissProt: Q9EQH3 Mouse
Unigene: 454528 Human
Unigene: 296520 Mouse
Unigene: 391632 Mouse
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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 kDa
Detected by chemiluminescence.
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).
Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (ab10099)This image is courtesy of an anonymous Abreview
Immunofluorescence 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VPS35 antibody (ab10099)
Negative Control showing staining of paraffin embedded Human Prostate, with no primary antibody.
