Anti-GAPDH antibody - Loading Control

• Supplier: Abcam
• Catalog: ab9485
• Prices: $403.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: IP ELISA WB IHC-F ICC/IF ICC/IF FC
• Species Reactivities: Mouse, Rat, Chicken, Dog, Human, Yeast, Xenopus, S. pombe, Monkey
• Antigen: Full length native protein from human erythrocytes
• Description: Rabbit Polyclonal
• Gene: GAPDH
• Conjugate: Unconjugated

Product images

ab9485 staining GAPDH in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab9485 at 5μg/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Rabbit secondary (ab150081) at 2 μg/ml (shown in green) and AlexaFluor®594 Goat anti-Mouse secondary (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

All lanes : Anti-GAPDH antibody - Loading Control (ab9485) at 1/2500 dilutionLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell LysateLane 3 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell LysateLysates/proteins at 20 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) at 1/10000 dilution

All lanes : Anti-GAPDH antibody - Loading Control (ab9485) 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 LysateLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

ab9485 staining GAPDH (green) in Rat bone marrow cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.1% Triton X 100 and blocked with 5% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/250 in PBS) for 12 hours at 4°C. An Alexa Fluor®488-conjugated goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody. Nuclei were stained with DAPI.See Abreview

ICC/IF image of ab9485 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab9485, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and 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).

All lanes : Anti-GAPDH antibody - Loading Control (ab9485) at 1/2500 dilutionLane 1 : Lysate prepared from human Huh-7 cells at 2 µgLane 2 : Lysate prepared from human Huh-7 cells at 20 µgSecondaryHRP-conjugated sheep polyclonal to rabbit IgG at 1/20000 dilutionPerformed under reducing conditions.

Product References

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