Products:Signal Transduction >> Signaling Pathway >> Nuclear Signaling >> NFkB Pathway
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Either no signal in WB, or seeing band of interest, plus a non-specific band. |
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ANSWER: |
Thank you for calling Abcam. |
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What is the immunogen sequence for this product? |
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ANSWER: |
Thank you for your enquiry. The immunogen sequence is near the N-terminus, in the general range 15 to 40 aa. The actual sequence information is proprietary and cannot be released but we can say that it is less than 25 aa long. |
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Deos this antibody require antigen retrieval? |
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ANSWER: |
Thank you for your enquiry. Further to correspondence with the source of this antibody I can tell you that we recommend the use of antigen retrieval for the application of this antibody bu IHC. I would recommend a standard citrate buffer approach. I hope this information helps, please do not hesitate to contact us if you need any more advice or information. |
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I just want to know the concentrations of ab32518 because there is no information on product datasheet for ab32518. Thank you very much in advance. |
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ANSWER: |
The IgG concentration for the current lot is 0.0427 mg/ml. Please contact us again if you have any additional questions.
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Anti-IKB alpha antibody [E130] (ab32518) at 1/10000 dilution + HeLa cell lysate
Predicted band size : 36 kDa
Observed band size : 35 kDa (why is the actual band size different from the predicted?)
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using ab32518 at 1/50 dilution.
ab32518 used to immunoprecipitate IKB alpha from human HeLa whole cell lysate. The antibody was further used to Western blot the protein.
Lane 1 IKB alpha IP
Lane 2 Control immunoprecipitate
Lane 3 Input (20%)
This image is courtesy of an anonymous Abreview
ab32518 used to immunoprecipitate IKB alpha from rat PC12 whole cell lysate. The antibody was further used to Western blot the protein.
Lane 1 IKB alpha IP
Lane 2 Control immunoprecipitate
Lane 3 Input (20%)
This image is courtesy of an anonymous Abreview
ab32518 at 1/100 staining mouse kidney tissue sections by IHC-P. The tissue was paraformaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed before the tissue was blocked and incubated with the antibody for 1 hour. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
This image is courtesy of an anonymous Abreview
ICC/IF image of ab32518 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 (ab32518, 1/1000 dilution) overnight at +4ºC. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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.
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