Anti-VPS26 抗体 (ab23892)
製品の概要
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製品名Anti-VPS26 antibody
VPS26 一次抗体 製品一覧 -
製品の詳細Rabbit polyclonal to VPS26
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由来種Rabbit
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アプリケーション適用あり: WB, IHC-P, ICC/IF, IPmore details
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種交差性交差種: Mouse, Rat, Human
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免疫原
Synthetic peptide conjugated to KLH derived from within residues 300 to the C-terminus of Human VPS26.
Immunogen の所有権に関して (Peptide available as ab24288.) -
ポジティブ･コントロール
- Human and mouse kidney lysate.
製品の特性
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製品の状態Liquid
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保存方法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.
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バッファーPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
精製度Immunogen affinity purified
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ポリ/モノポリクローナル
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アイソタイプIgG
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研究分野
関連製品
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Alternative Products
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Compatible Secondaries
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Isotype control
アプリケーション
Our Abpromise guarantee covers the use of ab23892 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| WB | Use a concentration of 1 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa). Abcam recommends using milk as the blocking agent. |
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| IHC-P | 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
| ICC/IF | Use a concentration of 1 µg/ml. | |
| IP | Use at 1 µg/mg of lysate. |
ターゲット情報
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機能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).
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配列類似性Belongs to the VPS26 family.
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細胞内局在Cytoplasm. Endosome membrane.
- Information by UniProt
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参照データベース
- Entrez Gene: 9559 Human
- Entrez Gene: 30930 Mouse
- Entrez Gene: 361846 Rat
- Omim: 605506 Human
- SwissProt: O75436 Human
- SwissProt: P40336 Mouse
- SwissProt: Q6AY86 Rat
- Unigene: 499925 Human
see all -
別名
- FLJ12930 antibody
- HB 58 antibody
- HB58 antibody
see all
画像
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Western blot - Anti-VPS26 antibody (ab23892)Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: VPS26 knockout HAP1 cell lysate (20 µg)
Lane 3: Jurkat cell lysate (20 µg)
Lanes 1 - 3: Merged signal (red and green). Green - ab23892 observed at 40 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab23892 was shown to recognize VPS26 when VPS26 knockout samples were used, along with additional cross-reactive bands. Wild-type and VPS26 knockout samples were subjected to SDS-PAGE. ab23892 at a concentration of 1 µg/ml and ab8245 (loading control to GAPDH) diluted to 1/10000 were incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VPS26 antibody (ab23892)Image courtesy of Human Protein Atlas
ab23892 staining VPS26 in Human kidney. The paraffin embedded tissue was incubated with ab23892 (1/500 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab23892 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines
Further results for this antibody can be found at www.proteinatlas.org
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Western blot - Anti-VPS26 antibody (ab23892)All lanes : Anti-VPS26 antibody (ab23892) at 1 µg/ml
Lane 1 : Human kidney lysate
Lane 2 : Human kidney lysate with Human VPS26 peptide (ab24288) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Alexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 40 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?
Additional bands at: 25 kDa, 50 kDa. We are unsure as to the identity of these extra bands.ab23892 detects a band of ~ 38kDa in Human kidney lysate. This band is quenched by the addition of the immunizing peptide, ab24288. ab23892 also detects a band of the correct size in HEK293 and mouse kidney lysate (data not shown).
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Immunocytochemistry/ Immunofluorescence - Anti-VPS26 antibody (ab23892)ICC/IF image of ab23892 stained human HEK 293 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab23892, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa and HepG2 cells.
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Western blot - Anti-VPS26 antibody (ab23892)This image is courtesy of an anonymous AbreviewAll lanes : Anti-VPS26 antibody (ab23892) at 1/1000 dilution
Lane 1 : Rat NRK whole cell lysate at 25 µg
Lane 2 : Rat NRK whole cell lysate at 50 µg
Secondary
All lanes : HRP-conjugated goat anti-rabbit polyclonal IgG at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 40 kDa
Observed band size: 40 kDa
Exposure time: 10 seconds
Blocked with 5% milk for 30 min at 25°C -
Immunocytochemistry/ Immunofluorescence - Anti-VPS26 antibody (ab23892)Image from Xia B et al., PLoS One. 2010 Jul 23;5(7):e11771. Fig 3.; doi:10.1371/journal.pone.0011771; July 23, 2010, PLoS ONE 5(7): e11771.Immunofluorescence analysis of HeLa cells treated with mDpy-30 siRNA, staining VPS26 (green) with ab23892.
Cells were fixed formaldehyde, permeabilized and blocked with 5% goat serum for 20 minutes. Samples were incubated with primary antibody in blocking buffer for 2 hours before incubating with fluorophore-conjugated secondary antibody for 1 hour. Nuclei were detected using DAPI. -
Western blot - Anti-VPS26 antibody (ab23892)This image is courtesy of an anonymous AbreviewAll lanes : Anti-VPS26 antibody (ab23892) at 1/1000 dilution
Lane 1 : HEK293T whole cell lysate
Lane 2 : HeLa whole cell lysate
Lysates/proteins at 50 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit polyclonal IgG at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 40 kDa
Observed band size: 40 kDa
Exposure time: 2 seconds
Blocked with 5% milk for 30 min at 25°C -
Immunocytochemistry/ Immunofluorescence - Anti-VPS26 antibody (ab23892)This image is courtesy of an anonymous Abreview.Immunocytochemical analysis of Rat NRK cell line, labeling VPS26 with ab23892. Cells were formaldehyde fixed, permeabilized with 0.1% Triton X-100 in PBS for 3 minutes and blocked with 1% BSA for 30 minutes at 25°C. Cells were incubated with ab23892 diluted 1/250 in 1% BSA in PBS for 1 hour at 25°C.
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Western blot - Anti-VPS26 antibody (ab23892)This image is courtesy of an anonymous Abreview.Anti-VPS26 antibody (ab23892) at 1/1000 dilution + COS whole cell lysate, African Green Monkey at 50 µg with Milk, 30 minutes, 25°C at 5 %
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 40 kDa
Exposure time: 10 seconds
プロトコール
データシートおよび資料
参考文献
This product has been referenced in:
- Toh WH et al. GGA1 regulates signal-dependent sorting of BACE1 to recycling endosomes, which moderates Aß production. Mol Biol Cell 29:191-208 (2018). Read more (PubMed: 29142073) »
- Seaman MNJ et al. Inhibition of TBC1D5 activates Rab7a and can enhance the function of the retromer cargo-selective complex. J Cell Sci 131:N/A (2018). Read more (PubMed: 29777037) »
