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ab25700 |
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I would use this antibody for rat tissue (IHC-P). However in the Abcam catalog, another antibody is suggested for the same application (i.e.Ab23375). Could you suggest me which would be the best antibody for this application? Thank you very much |
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ANSWER: |
Thank you for your enquiry and your interest in our products.
I can confirm that both antibodies recognize rat ALDH1A1, however the immunogens used to raise these antibodies were different. Whilst, for ab23375 a synthetic peptide conjugated to KLH derived from within residues 450 to the C-terminus of - rat - ALDH1A1 was used; for the other antibody ab52492, the immunogen peptide corresponding to residues near the C-terminus of - human - Aldh1a1 was applied.
I hope this helps and if I can assist further, please do not hesitate to contact me. |
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
ICC/IF image of ab23375 stained rat PC12 cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab23375, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).
IHC image of ALDH1A1 staining in rat brain FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab23375, 1µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
ab23375 staining ALDH1A1 in Mouse hypothalamic tanycytes by IHC-FrFl (Immunohistochemistry Free-floating). 40 µm free-floating sections were incubated with primary antibody (1/300) for 18 hours at 4ºC in blocking solution. An Alexa Fluor® 555-conjugated Donkey polyclonal to rabbit IgG (1/300) was used as secondary antibody.
This image is courtesy of an anonymous Abreview
All lanes : Anti-ALDH1A1 antibody - Neuronal Marker (ab23375) at 1 µg/ml
Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 2 : Liver (Mouse) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 54 kDa
Observed band size : 54 kDa
Additional bands at : 95 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 90 seconds
All lanes : Anti-ALDH1A1 antibody - Neuronal Marker (ab23375) at 1 µg/ml
Lane 1 : Brain (Mouse) Tissue Lysate
Lane 2 : Liver (Mouse) Tissue Lysate
Lane 3 : Liver (Rat) Tissue Lysate
Lane 4 : E14 Mouse Embryo Brain Tissue Lysate
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 54 kDa
Observed band size : 51,54 kDa (why is the actual band size different from the predicted?)
Exposure time : 90 seconds
Anti-ALDH1A1 antibody - Neuronal Marker (ab23375) at 1 µg/ml +
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Exposure time : 1 minute
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