Anti-beta IV Tubulin antibody [ONS.1A6]

• Supplier: Abcam
• Catalog: ab11315
• Prices: $390.00
• Sizes: 100 µg
• Host: Mouse
• Clonality: Monoclonal
• Isotype: IgG1
• Clone: ONS.1A6
• Applications: IHC-P ICC/IF ICC/IF IHC-F ELISA WB
• Species Reactivities: Mouse, Human, Rat, Chicken, Hamster, Bovine
• Antigen: Synthetic peptide corresponding to the C-terminal sequence, coupled to BSA
• Description: Mouse Monoclonal
• Gene: TUBB4A
• Conjugate: Unconjugated

Product images

All lanes : Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315) at 5 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear LysateLane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell LysateLane 4 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell LysateLane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 6 : U2OS (Human osteosarcoma cell line) Whole Cell LysateLysates/proteins at 20 µg per lane.SecondaryGoat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

IHC image of beta IV Tubulin staining in Human normal skin 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 ab11315, 1µg/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. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

ICC/IF image of ab11315 stained MCF-7 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 ab11315 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) 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.

Immunofluorescence analysis of MCF-7 Cells (human breast cancer cell line), staining beta IV Tubulin with ab11315. Cells on coverslip were fixed with -20°C Methanol for 5 min before permeabilization with 0.2% Triton X-100 in PBS for 10 min. Cells were incubated with the primary antibody with a dilution 1/50 for 1 hour. After brief washing with PBS, cells were incubated with TRITC conjugated secondary antibody with a dilution 1/50 for 45 min and washed with PBS to be examined under fluorescence microscope.

Immunofluorescence analysis of MCF-7 Cells (human breast cancer cell line), staining beta IV Tubulin with ab11315. Cells on coverslip were fixed with -20°C Methanol for 5 min before permeabilization with 0.2% Triton X-100 in PBS for 10 min. Cells were incubated with the primary antibody with a dilution 1/50 for 1 hour. After brief washing with PBS, cells were incubated with TRITC conjugated secondary antibody with a dilution 1/50 for 45 min and washed with PBS to be examined under fluorescence microscope.

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