The antibody produces a strong postive labeling of LHRH / GnRH at dilution of 1/2,000 - 1,4,000 using biotin-streptavidin/HRP in rat hypothalamus (median eminence). Optimal dilution will vary depending upon fixation, labeling technique and/or detection system; therefore, a dilution series is recommended.
The above image is of LHRH (GnRH) terminals in rat median eminence, ryostat sections, ABC / DAB detection.
Staining is completely eliminated by pretreatment of the diluted antibody with 5 µg of LHRH per mL of diluted antiserum.
Photo Description: IHC image of staining for LNRH/GnRH in the rat median eminence. The tissue was fixed with 4% formaldehyde in phosphate buffer, before being removed and prepared for vibratome sectioning. Floating sections were incubated at RT in 10% goat serum in PBS, before standard IHC procedure. Primary antibody was incubated at 1:4000 for 48 hours, goat anti-rabbit secondary was subsequently added for 1 hour after washing with PBS. Light microscopy staining was achieved with standard biotin-streptavidin/HRP procedure and DAB chromogen.
Quantity / Volume: 100 µL
State: Lyophilized Whole Serum
Reacts With: Bat, Carp, Catfish, Chick, Chicken, Clarias Batrachus (Catfish), Fish, Frog, Frog (Green Treefrog), Gerbil, Guinea Pig, Guinea Pig, Hagfish, Hamster, Human, Lizard, Mole Rat, Monkey, Mouse, Mudpuppy (Necturus Maculosus), Octopus, Pig, Rat, Salamander, Snail, Snail (Bulla Gouldiana), Squid, Trout
Availability: In Stock
Alternate Names: Gonadotropin-Releasing Hormone; Gonadorelin; Luliberin I; Gonadoliberin I; progonadoliberin-1; GRH, anti-LHRH
Gene Symbol / ID, Accession #: GNRH1, 2796
Highly recommend -- very specific staining with no background
I cut 10-20 microns sections, dry them and the process them as following: Protocol: 1hr blocking (PBS/0.1% triton plus 10% NGS), incubate overnight with LHRH antibody diluted in PBS/0.1% triton, 1:400. Then washes and secondary antibody fluorescent. When used with secondary biotinylated antibody, I use LHRH antibody less diluted (1:4000).
My student is also using the LHRH antibody on paraffin sections, with the same results (and also in conjunction with Tyramide amplification kit).
Staining is very specific. Stronger on adult tissue because more GnRH expressed by the neurons producing it. There is no background. Fixation sensitive; long fixations are not recommended. I highly recommend this antibody.
Anna Cariboni, PhD
Department of Cell and Developmental Biology
University College London
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Q & A Wall
I am interested in using LHRH antibody (Cat# 20075) for fixed cells ELISA assay. Do you recommend biotin-adivin/HRP system because it worked best for this antibody or you have not really tried anti rabbit IgG-HRP detection system?
Question By Dina
I see no reason why it wouldn't work. The use of anti-rabbit HRP is okay; they just need to work out an optimal concentration of LHRH aby and the secondary. I assume they will fix with formalin too...sometime cells are fixes in acetone which preserves some, but not all, antigens for immunohistochemistry so formalin is recommended.
Answer By Colleen on 2015-11-20 13:43:26
This product contains the preservative sodium azide. The concentration percent of the sodium azide is ≤ .09%. Although this hazardous substance is a concentration below that required for the preparation of a Material Safety Data Sheet, we created a standard MSDS for your records.Download MSDS