Anti-Glucose Transporter GLUT1抗体
参阅全部 Glucose Transporter GLUT1 一抗
兔多克隆抗体to Glucose Transporter GLUT1
Rabbit
适用于: IHC-P, ICC/IF, WBmore details
与反应: Human
预测可用于: Horse, Chicken, Cat, Pig, Chimpanzee, Monkey, Baboon
Synthetic peptide corresponding to Rat Glucose Transporter GLUT1 aa 481-492 (C terminal) (Cysteine residue).
Sequence:
[C]GLFHPLGADSQV
Database link: P11167
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Liquid
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 30% Glycerol (glycerin, glycerine), 0.5% BSA, 0.24% HEPES, 0.16% Tris HCl
浓度
批次浓度范围 50 µg 浓度为 0.5 - 0.78 mg/ml
Immunogen affinity purified
Purified antibody was isolated from immobilized antigen sepharose affinity column.
多克隆
IgG
Abpromise™承诺保证使用ab14683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 at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
| ICC/IF | 1/200. | |
| WB | 1/2500. Detects a band of approximately 50 kDa (predicted molecular weight: 55 kDa). We suggest that samples should not be heated or frozen. |
Entrez Gene: 396130 Chicken
Entrez Gene: 6513 Human
Omim: 138140 Human
SwissProt: P46896 Chicken
SwissProt: P11166 Human
Unigene: 473721 Human
Unigene: 721551 Human
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Western blot - Anti-Glucose Transporter GLUT1 antibody (ab14683)
All lanes : Anti-Glucose Transporter GLUT1 antibody (ab14683) at 1/2500 dilution
Lane 1 : Human placenta tissue lysate - total protein (ab29745)
Lane 2 : SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Predicted band size: 55 kDa
Observed band size: 55 kDa
This antibody labels Glut 1 protein as a smear due to large amount of glycosylation on Glut 1 transporter. We observed that boiling and freezing the Glut 1 samples resulted in aggregated form of the protein and gives a smear form high 100kDa range to 45kDa with significant reduction in Glut 1 (44-47kDa) band. We suggest that samples should not be heated or frozen. Following extraction in lysis buffer, the proteins are then dissolved in SDS-PAGE sample buffer 2X and reduced in presence of 2.5-5% BME. All samples at this time will be heated 70-80°C for 3 minutes and aliquot and stored frozen till it is time for analysis. Just before analyses the samples much be thawed at room temp or at 37°C to dissolve any precipitated SDS in the sample. The remaining sample must not be frozen again, this freeze and next thaw cycle will make aggregate in some samples.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody (ab14683)
Human normal placenta. Staining is observed at the cell surface.
Left panel: Primary antibody at 2 ug/ml.
Right panel: Isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH 6.1 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutess. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako envision flex amplification kit for rabbit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with haematoxylin and coverslipped under DePeX.
Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
Immunocytochemistry/ Immunofluorescence - Anti-Glucose Transporter GLUT1 antibody (ab14683)
ICC/IF image of ab14683 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells. The cells were 4% formaldehyde fixed (10 minutes) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab14683, 5 µg/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 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody (ab14683)Image from Paatero I et al., J Biol Chem. 2012 Mar 23;287(13):9659-71. Epub 2012 Feb 3. Fig 1.; doi: 10.1074/jbc.M111.299537; March 23, 2012 The Journal of Biological Chemistry, 287, 9659-9671.
Immunohistochemical analysis of mammary glands taken from wild-type or Erbb4-/- lactating mice, staining Glucose Transporter GLUT1 with ab14683.
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