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Western blot analysis of PAX5 was performed by loading 75µg of whole cell lysates from PAX5-expressing B cell lymphoma-derived cell lines, negative control Jurkat cells, and 10ul of PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a PAX5 polyclonal antibody (Product # PA1-109) at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBS-0.1% Tween-20, and probed with a goat anti-rabbit HRP secondary antibody (Product # 32460) at a dilution of 1:500 (20ng/ml) for one hour. Membranes were washed and chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080, upper panel). To assay for protein loading (lower panel), the membrane was stripped with Restore PLUS Western Blot Stripping Buffer (Product # 46430), blocked with 5% BSA/TBST, and re-probed with an HRP-conjugated Beta Tubulin Loading Control Antibody (Product # MA5-16308-HRP) at a dilution of 1:1000 for 1 hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).
|Tested species reactivity||Rabbit|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with proprietary stabilizer|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:60-1:500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 2 publications below|
This antibody may cross-react with immunoglobulins from other species.
Working dilutions for Western blotting using SuperSignal West Substrates:
SuperSignal West Pico Substrate - 1:250-1:1250
SuperSignal West Dura Substrate - 1:600-1:3000
SuperSignal West Femto Substrate - 1:1250-1:6000
ECL Western Blotting Substrate - 1:20-1:125
Thermo Scientific Anti-Rabbit secondary antibodies are affinity-purified antibodies with well-characterized specificity for rabbit immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Regulation of the transcriptional activation of the androgen receptor by the UXT-binding protein VHL.
32460 was used in western blot to study the mechanism by which the UXT-binding protein VHL modulates androgen receptor transcriptional activation
|Chen S,Chen K,Zhang Q,Cheng H,Zhou R||The Biochemical journal (456:55)||2013|
|Not Applicable||Not Cited||
The SUMOylation of zinc-fingers and homeoboxes 1 (ZHX1) by Ubc9 regulates its stability and transcriptional repression activity.
32460 was used in western blot to study the mechanism by which Ubc9-mediated SUMOylation of ZHX1 modulates its stability and functional activity
|Chen S,Yu X,Lei Q,Ma L,Guo D||Journal of cellular biochemistry (114:2323)||2013|