Product nameAnti-Rhox5 / PEM antibody
DescriptionRabbit polyclonal to Rhox5 / PEM
Tested applicationsSuitable for: ICC/IF, IHC-P, WBmore details
Species reactivityReacts with: Mouse
- Mouse Embryonic Germ Cell Lysate This antibody gave a positive result in IHC in the following FFPE tissue: Mouse normal testis. IF/ICC: NIH 3T3 cell line.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab31922 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||Use a concentration of 1 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 30 kDa (predicted molecular weight: 23 kDa).|
RelevanceRhox5 / PEM is a putative transcription factor which is developmentally regulated. It is expressed in female mouse embryos only from their paternal X chromosome due to early epigenetic gene regulation. Rhox5 can be used to identify female mouse embryos after just 3.5 days.
- Homeobox protein Pem antibody
- Homeobox protein Rhox5 antibody
- PEM antibody
- Rhox5 antibody
ab31922 stained NIH 3T3 cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab31922 at 1µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
IHC image of Rhox5 / PEM staining in Mouse normal testis formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab31922, 1µg/ml, for 15 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.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
All lanes : Anti-Rhox5 / PEM antibody (ab31922) at 1/1000 dilution
Lane 1 : MSCI cells (mouse sertoli cell line) untransfected
Lane 2 : MSCI cells (mouse sertoli cell line) transfected with FLAG-tagged Rhox5
Lanes 3-4 : Mouse Rhox5 knockout testis
Lane 5 : Day 31 mouse testis
Lanes 6-7 : FLAG-Rhox5 transgenic mouse testis
Lysates/proteins at 5 µg per lane.
All lanes : GE Healthcare Sheep anti Mouse - HRP conjugated at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?
Additional bands at: 40 kDa (possible cross reactivity)
Exposure time: 30 seconds
This product has been referenced in:
- Hu X et al. Deletion of the tyrosine phosphatase Shp2 in Sertoli cells causes infertility in mice. Sci Rep 5:12982 (2015). Mouse . Read more (PubMed: 26265072) »
- Li Q et al. Three epigenetic drugs up-regulate homeobox gene Rhox5 in cancer cells through overlapping and distinct molecular mechanisms. Mol Pharmacol 76:1072-81 (2009). Read more (PubMed: 19679824) »