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Thanks, My question is ciontrol antibody ? What is the concentration of your Ab in 500 microL?
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ANSWER: |
You could use any unrelated antibody for a negative control, we don't have a specific one to recommend. You can also run a negative control using the same conditions that would be appropriate for antibody staining, replacing the primary antibody with blocking serum. Ab9514 is provided in the form of tissue culture supernatant, and since this is unpurified material, we are unable to provide an exact concentration. |
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What type of tissue is recommended as a negative control? |
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ANSWER: |
Thank you for your enquiry. There is no particular negative control tissue for CD74; the negative control should be run on the same tissue (known positive) but without primary antibody. 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"
ab9514 staining human tonsil by IHC-P.
Anti-CD74 antibody [LN2] (ab9514) at 5 µg/ml +
Secondary
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Predicted band size : 34 kDa
Observed band size : 34 kDa
Additional bands at : 37 kDa (possible post-translational modification).
ab9514 staining CD74 in human liver tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with neutral buffered formalin and a heat mediated antigen retrieval step was performed using EDTA buffer pH 9.0. Samples were then blocked with 10% serum for 10 minutes at 20ºC followed by incubation with the primary antibody at a 1/75 dilution for 30 minutes at 20ºC. A HRP-conjugated rat anti-mouse/rabbit polyclonal was used undiluted as the secondary antibody.
Image courtesy of an anonymous Abreview.
Overlay histogram showing Raji cells stained with ab9514 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9514, 1/10 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (
Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.
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