Products:Cell Biology >> Cell Cycle >> Cell Cycle Inhibitors >> Rb
If your product does not perform as described on this datasheet, we will refund or replace your product...
Read our guarantee »This product is covered by the Abpromise guarantee. Our scientific support team are available to answer any questions or queries - fill out an inquiry form for ab24 for help.
Alternatively, you can search the previous enquiries about this product to see if your query has already been answered.
|
|||||||||
|
|||||||||
I would like to know if this antibody works well as nuclear marker in rat lysates
|
|||||||||
ANSWER: |
Thank you for your enquiry. Ab24 is predicted to react with rat Rb (97% identity with immunogen) due to sequence homology but this has not been specifically tested yet. Should you decide to go ahead and purchase this product, please let us know how you get on by submitting an Abreview and in return we will award you 50 Abcam Points, which can be redeemed on a number of rewards (a further 100 points will be offered for an image). Please let me know if you require any further assistance.
|
||||||||
|
|||||||||
I would like to follow-up with your tech service department before ordering ab24 or ab3077. I am hoping that you can verify that either or both of these 2 pRB antibodies are useful for detection of the active form of RB. I am hoping to probe with these to determine if pRB is "turned on" and not just present. |
|||||||||
ANSWER: |
Thank you for your enquiry. Ab24 also recognises the phosphorylates and unphospohorylated forms of Rb1, so it is nonspecific. I hope this information helps, please do not hesitate to contact us if you need any more advice or information, |
||||||||
|
|||||||||
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Anti-Rb antibody [Rb1 1F8] - Nuclear Marker (ab24) at 1/1000 dilution + Human Fibroblast cell lysate(nuclear). 16 hour incubation.
Secondary
NIF 825 (provided in ECL kit) - HRP conjugated
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 105 kDa
Observed band size : 100-150 kDa (why is the actual band size different from the predicted?)
Exposure time : 1 minute
This image is courtesy of an Abreview submitted on 5 October 2005.
Predicted band size : 105 kDa
Lysates prepared from lung and kidney of VerUTR transgenic and wildtype mice were analyzed by Western Blot probed with ab24. Increased expression of Rb1 was detected in the organs from the VerUTR transgenic mice.
Cells were seeded onto 6-well plates at 2×105 cells per well overnight. They were then transfected with 1 µg of VerUTR or control vector in combination with scrambled RNA or siRNA against VerUTR. Proteins were extracted 48 hours after transfection by lysing in 60 µl of lysis buffer containing protease inhibitors (150 mM NaCl, 25 mM Tris-HCl, pH 8.0, 0.5 mM EDTA, 1% Triton X-100, 8 M Urea, and 1x protease inhibitor cocktail). Tissues were disrupted in appropriate volume of lysis buffer depending on tissue weight. All samples were subjected to SDS-PAGE and then transferred to nitrocellulose membranes followed by incubating with ab24 at 1/500 dilution at 4°C overnight. The secondary antibody used was goat anti-mouse IgG at 1/2000 dilution at room temperature for 1 hour. After detection of the protein bands, the blot was stripped and re-probed with mouse monoclonal antibody against β-actin to confirm equal loading. After secondary antibody incubation, the blot was washed and detected by ECL kit in autoradiography.
Image from Lee DY et al, PLoS One 5:e13599 (2010), Fig 3.
3
Call 01223 696 000 or contact us