Anti-GAPDH antibody [mAbcam 9484] - Loading Control

• Supplier: Abcam
• Catalog: ab9484
• Prices: $401.00
• Sizes: 200 µg
• Host: Mouse
• Clonality: Monoclonal
• Isotype: IgG2b
• Clone: mAbcam 9484
• Applications: ICC/IF ICC/IF WB IHC-P FC
• Species Reactivities: Mouse, Rat, Rabbit, Chicken, Bovine, Dog, Human, Pig, Xenopus, Monkey, Hamster
• Antigen: Full length native protein from human erythrocytes
• Description: Mouse Monoclonal
• Gene: GAPDH
• Conjugate: Unconjugated

Product images

All lanes : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/5000 dilutionLane 1 : Hela whole cell (Human)Lane 2 : 3T3 cell (Mouse)Lane 3 : Rat brainLane 4 : Xenopus embryoLane 5 : Chicken LiverLane 6 : EBTr cell (Cow)Lane 7 : CHO cell (Chinese hamster)Lane 8 : Pig liverSecondaryRabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilutionPerformed under reducing conditions.

Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 0.5 µg/ml + HeLa cell lysateSecondaryGoat Anti-Mouse IgG H&L (HRP) (ab6789) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under non-reducing conditions.

Lanes 1 - 5 : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/1000 dilution (Blocked in 5% MILK)Lanes 6 - 10 : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/1000 dilution (Blocked in 5% BSA)Lane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear LysateLane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)Lane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate (ab7902)Lane 6 : HeLa (Human epithelial carcinoma cell line) Nuclear LysateLane 7 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 8 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)Lane 9 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)Lane 10 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate (ab7902)Lysates/protein

IHC image of GAPDH staining in human liver FFPE section, performed on a 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 ab9484, 5µg/ml, for 8 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 ab9484 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab9484, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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.

ICC/IF image of ab9484 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab9484, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, anti mouse-DyLight® 488 (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.

Flow Cytometry analysis of GAPDH using ab9484.Human monocytes were fixed in paraformaldehyde and permeabilized. ab9484 was used at a 1/200 dilution. The secondary used was an Alexa-Fluor 488 conjugated chicken anti-rabbit IgG (H+L) polyclonal, used at a 1/500 dilution.See Abreview

Overlay histogram showing HeLa cells stained with ab9484 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab9484, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was goat anti-mouse DyLight® 488 (IgG H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

Product References

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