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Western blot
ELISA
ImmunohistochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
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- Product number
- LS-B52 - Provider product page
- Provider
- LSBio
- Product name
- IHC-plus™ E2F1 Antibody (phospho-Ser364) LS-B52
- Antibody type
- Polyclonal
- Description
- Affinity purified
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Storage
- Store at 4°C or -20°C. Avoid freeze-thaw cycles.
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Supportive validation
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- LSBio (provider)
- Enhanced method
- Genetic validation
- Main image
- Experimental details
- Western blot of Affinity Purified anti-E2F-1 pS364 antibody shows detection of a band at ~47 kD corresponding to phosphorylated E2F-1 in induced cell lysates. Panel A shows reactivity using a control antibody reactive to all forms of E2F (arrowheads). Panel B shows specific reactivity against phosphorylated E2F-1 (arrowheads) using our anti-E2F-1 pS364 antibody. Lysates are as follows: CRE/E2F-1 are CRE cells derived from mouse NIH3T3 line transfected with human E2F-1, NIH-3T3 used as a negative control, and MDA-MB-231 cells are a human breast cancer line. As indicated each lysate was prepared from untreated cells and cells treated with 2 uM Doxorubicin used as a DNA damaging agent. In addition the MDA-MB-231 cells were also treated with genistein, a mild DNA damaging agent. The figure shows the same membrane first probed with the anti-E2F-1 pS364 antibody used at a 1:250 dilution, then stripped and re-probed with the pan E2F antibody used as a positive control. The positive control antibody clearly shows an E2F-1 band in all human cell lines, but not mouse cells. Treatment with doxorubicin increases expression of E2F-1 as shown in Panel A. Images were overlapped to confirm that anti-E2F-1 pS364 staining shown treated human cells in Panel B specifically aligns with E2F-1 staining shown in Panel A. Blots can be processed with HRP conjugated Gt-a-Rabbit IgG MX10 LS-C60865 for 45 min at room temperature for ECL detection. Personal Communication, XiaoHe Yang, Univ. Oklahoma.
