Anti-Glucose Transporter GLUT1 antibody

• Supplier: Abcam
• Catalog: ab32551
• Prices: $390.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: ICC/IF ICC/IF WB IHC-P IP
• Species Reactivities: Mouse, Rat, Human, Rabbit
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Glucose Transporter GLUT1
• Description: Rabbit Polyclonal
• Gene: SLC2A1
• Conjugate: Unconjugated

Product images

All lanes : Anti-Glucose Transporter GLUT1 antibody (ab32551) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate (ab7902)Lysates/proteins at 10 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilutionPerformed under reducing conditions.

All lanes : Anti-Glucose Transporter GLUT1 antibody (ab32551) at 1 µg/mlLane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate Lane 3 : Liver (Mouse) Tissue Lysate - normal tissue Lane 4 : Kidney (Mouse) Tissue Lysate Lane 5 : Testis (Mouse) Tissue Lysate - normal tissue Lane 6 : Spinal Cord (Mouse) Tissue Lysate Lane 7 : Ovary (Mouse) Tissue Lysate - normal tissue Lane 8 : Liver (Rat) Tissue Lysate Lysates/proteins at 10 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilutionPerformed under reducing conditions.

IHC image of Glucose Transporter GLUT1 staining in human kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32551, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

ICC/IF image of ab32551 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32551, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Glucose Transporter GLUT1 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to Glucose Transporter GLUT1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32551.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 51kDa; non specific band - 37kDa: We are unsure as to the identity of this extra band. Glucose Transporter GLUT1

Product References

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