Products:Tags & Cell Markers >> Subcellular Markers >> Organelles >> Mitochondria
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ab16131 |
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ab16131 |
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Would like to stain mitochondria in NIH3T3 cells. |
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ANSWER: |
Thank you for your call today and for your enquiry. |
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We are thinking of buying two antibodies ab15895, ab69090 from abcam. What we are trying to do is, measure the amount of ferritin and mitochondria at C57bl6 mouse brain cells. Please let us know whether these two antibodies will work or not? |
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ANSWER: |
After looking at these antibodies I think that they will most likely be appropriate for your use; however it does depend on the type of experiment you are performing. Both of these antibodies are tested and guaranteed to work in immunocytochemistry, immunohistochemistry and western blotting. If you are not planning on using the antibodies in one of these applications I would be happy to find an antibody appropriate for the application that you will be using. |
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
All lanes : Anti-VDAC1 / Porin antibody - Mitochondrial Loading Control (ab15895) at 1 µg/ml
Lane 1 : Hela Nuclear lysate
Lane 2 : Hela cell lysate
Lane 3 : A431 cell lysate
Lane 4 : Jurkat cell lysate
Lane 5 : HEK293 cell lysate
Lane 6 : Hela Nuclear lysate with
Lane 7 : Hela cell lysate with
Lane 8 : A431 cell lysate with
Lane 9 : Jurkat cell lysate with
Lane 10 : HEK293 cell lysate with
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 31 kDa
Observed band size : 31 kDa
ICC/IF image of ab15895 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab15895, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 phalloidin was used to label F-actin (red).
Image courtesy of Human Protein Atlas
Paraffin embedded sections of human kidney were incubated with ab15895 (1/600 dilution) at room temperature for 30 mins. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab15895 was also tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further images can be found at www.proteinatlas.org
All lanes : Anti-VDAC1 / Porin antibody - Mitochondrial Loading Control (ab15895) at 1 µg/ml
Lane 1 : Heart (Mouse) Tissue Lysate
Lane 2 : Kidney (Mouse) Tissue Lysate
Lane 3 :
Lane 4 : Spinal Cord (Mouse) Tissue Lysate
Lane 5 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lane 6 : Brain (Rat) Tissue Lysate - normal tissue
Lane 7 :
Lysates/proteins at 10 µg per lane.
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 31 kDa
Observed band size : 31 kDa
ab15895 staining mouse heart tissue section by IHC-P. Sections were PFA fixed and subjected to heat mediated antigen retrieval in citrate buffer (pH 6) prior to blocking in 10% serum for 10 minutes at 25°C. The primary antibody was diluted 1/600 and incubated with the sample for 1 hour at 25°C. A biotinylated goat anti-rabbit antibody diluted 1/400 was used as the secondary.
This image is courtesy of an anonymous Abreview
All lanes : Anti-VDAC1 / Porin antibody - Mitochondrial Loading Control (ab15895) at 1 µg/ml
Lane 1 : Adult wild type zebrafish brain mitochondrial lysate at 20 µg
Lane 2 : Adult wild type zebrafish brain cytosolic lysate at 30 µg
Secondary
HRP-conjugated goat F(ab’)2 Fragment at 1/10000 dilution
developed using the ECL technique
Predicted band size : 31 kDa
Observed band size : 28,29 kDa (why is the actual band size different from the predicted?)
Exposure time : 1 minute
Image courtesy of an anonymous Abreview.
ab15895 staining VDAC1 / Porin in human cerebellum tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).Tissue was fixed in paraformaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer pH 6.0. Samples were then incubated with ab15895 at a 1/1000 dilution for 12 hours at 4°C. The secondary used was an undiluted HRP conjugated anti-rabbit polyclonal. Note: The big cells are purkinje cells of the cerebellum, (no nuclear staining).
Image courtesy of Dr Markus Kipp by Abreview.
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 31 kDa
Exposure time : 3 minutes
Western blot image using the Optiblot Reducing Electrophoresis Kit - 10 x 10 cm (4-20%) (ab119220) with the
Prism Ultra Protein Ladder (ab116028) 5µl used. We recommend using our ECL substrate kit (ab65623) .
20ug of Lysate per lane and detection using ab15895 diluted to 1ug/ml.
Lane 1: Hela cell lysate
Lane 2: Jurkat cell lysate
Lane 3: A431 cell lysate
Lane 4: HEK293 cell lysate
Lane 5: HepG2 cell lysate.
All lanes : Anti-VDAC1 / Porin antibody - Mitochondrial Loading Control (ab15895) at 1 µg/ml
Lane 1 : Chicken liver cell lysate
Lane 2 : CHO K1 cell lysate
Lane 3 : MDCK cell lysate
Lane 4 : Chicken liver cell lysate with
Lane 5 : CHO K1 cell lysate with
Lane 6 : MDCK cell lysate with
Lysates/proteins at 20 µg per lane.
Secondary
Alexa fluor goat polyclonal anti-Rabbit IgG at 1/10000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 31 kDa
Observed band size : 31 kDa
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