Anti stra8

Manufactured by Abcam

Sourced in United States, Germany

Anti-STRA8 is a primary antibody used to detect the STRA8 (Stimulated by Retinoic Acid gene 8) protein. STRA8 is a key regulator involved in meiosis initiation during gametogenesis. The Anti-STRA8 antibody can be used for applications such as western blotting, immunohistochemistry, and immunocytochemistry to study the expression and localization of the STRA8 protein.

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Lab products found in correlation

Vectashield medium with dapi, by Vector Laboratories (1 mentions) Texas red or fitc conjugated secondary antibodies, by Jackson ImmunoResearch (1 mentions) Abc reagent, by Vector Laboratories (1 mentions) Dab peroxidase substrate, by Vector Laboratories (1 mentions) Normal horse serum, by Vector Laboratories (1 mentions) Dab substrate kit, by Vector Laboratories (1 mentions) Permount, by Thermo Fisher Scientific (1 mentions) Anti dmc1, by Santa Cruz Biotechnology (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Axio microscope, by Zeiss (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Paraformaldehyde (pfa), by Merck Group (1 mentions) Alexa fluor 568 conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 568 conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Anti γh2ax, by Abcam (1 mentions) Dapi fluoromount g, by Southern Biotech (1 mentions) Anti sox9 sry box transcription factor 9, by Merck Group (1 mentions) Anti alpha smooth muscle actin, by Abcam (1 mentions) Sp5 confocal microscope, by Leica (1 mentions) Cy5 conjugated anti rabbit, by Jackson ImmunoResearch (1 mentions)

Spelling variants of this product

Stra8 (8 citations) Rabbit anti-STRA8 antibody (2 citations)

4 protocols using anti stra8

Fetal Gonad Characterization Immunohistochemistry

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Fetal gonads were dissected in phosphate buffered saline (PBS), fixed in 4% paraformaldehyde overnight at 4°C, embedded in paraffin and sectioned. Slides were incubated with anti-GCNA IgM (courtesy of G. Enders, undiluted supernatant), anti-STRA8 (Abcam. 1∶100), and anti-phosphoH2A.X (Upstate Cell Signaling Solutions, 1∶250 dilution). Colorimetric staining was performed using ABC reagents (Vector Laboratories) and developed with DAB peroxidase substrate (Vector Laboratories).
Sections were mounted in Vectashield Medium with DAPI (Vector Laboratories), and fluorescent staining was obtained using Texas-Red or FITC-conjugated secondary antibodies (Jackson Immunoresearch Laboratories, 1∶500 dilution).

Koubova J., Hu Y.C., Bhattacharyya T., Soh Y.Q., Gill M.E., Goodheart M.L., Hogarth C.A., Griswold M.D, & Page D.C. (2014). Retinoic Acid Activates Two Pathways Required for Meiosis in Mice. PLoS Genetics, 10(8), e1004541.

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Immunohistochemical Analysis of Spermatogenic Markers

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Adult testes were fixed in Bouin's solution overnight at 4°C, washed with PBS and 70% ethanol, embedded in paraffin, and sectioned at 5 µm thickness. Slides were matured overnight, de-waxed, rehydrated, and heated in 10 mM sodium citrate buffer (pH 6.0) for antigen retrieval. Sections were incubated in 3% hydrogen peroxide for 5 min and blocked in 2.5% normal horse serum (Vector Laboratories) for 80 minutes at room temperature. Later, slides were incubated overnight with anti-STRA8 (Abcam, 1∶500) or anti-DMC1 (Santa Cruz Biotechnology, 1∶50 dilution). The following day, slides were washed three times in PBS and incubated with anti-rabbit ImmPRESS peroxidase reagent (Vector Laboratories) for 30 minutes. The slides were later developed using a DAB substrate kit (Vector Laboratories) for 1 minute. The slides were counterstained with Mayer's hematoxylin for 5 minutes and washed in running water, dehydrated, and mounted with Permount (Fisher Scientific).

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Immunostaining of Germ Cells

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After 24 or 48h of culture, morphologically healthy germ cells (about 200–400 cells per set) were placed on glass slides and fixed with 4% paraformaldehyde (Sigma-Aldrich) for 15 min at room temperature. After washing, the cells were permeabilized with 0.2% Triton X-100 in PBS for 15 min, then blocked with 5% bovine serum albumin (BSA; Sigma-Aldrich) in PBS for 1h at room temperature. These cells were incubated with primary antibodies overnight at 4°C. Primary antibodies for immunostaining included (S1 Table): rabbit polyclonal anti-STRA8 (1:500; Abcam, Cambridge, MA, USA) and mouse polyclonal anti-γH2AX (1:500; Abcam). After washing, the cells were incubated with Alexa Fluor 568-conjugated goat anti-rabbit IgG (1:500; Thermo Fisher Scientific, Waltham, MA, USA) or Alexa Fluor 568-conjugated goat anti-mouse IgG (1:500; Thermo Fisher Scientific) (S1 Table) in 1% BSA in PBS, and subsequently counterstained with Hoechst 33342 (Invitrogen) for 15 min and mounted in 50% (v/v) glycerol in PBS. STRA8-positive and γH2AX-positive cells were detected and imaged using an Axio microscope (Carl Zeiss Microimaging, Göttingen, Germany). In the present study, the immunochemical staining was repeated independently in triplicate for each experiment.

Kim S.M., Yokoyama T., Ng D., Ulu F, & Yamazaki Y. (2019). Retinoic acid-stimulated ERK1/2 pathway regulates meiotic initiation in cultured fetal germ cells. PLoS ONE, 14(11), e0224628.

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Immunohistochemical Analysis of Mouse Testes

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Adult mice were anesthetized with isoflurane and transcardially perfused with PBS, followed by 4% PFA. Testes were dissected and post-fixed in 4% PFA overnight. Testis sections were blocked (20% horse serum in PBS, one hour, room temperature) and incubated with primary antibodies in PBS containing 0.3% Triton X-100 and 10% horse serum (overnight, 4 °C). Primary antibodies used were sheep polyclonal anti-Np detecting Np65 and Np55 (pan-Np55/65; 1:300, R&D systems) and rabbit polyclonal anti-Stra8 (Stimulated By Retinoic Acid 8, used as a marker for spermatogonia and spermatocytes, 1:500, Abcam), anti-CPY11A1 (Cytochrome P450 Family 11 Subfamily A Member 1, marker for Leydig cells, 1:500, Abcam), anti-alpha-smooth muscle actin (marker for myoid cells, 1:500, Abcam), and anti-Sox9 (SRY-Box Transcription Factor 9, marker for Sertoli cells, 1:500, EMD Millipore, Darmstadt, Germany) antibodies. The specificity of antibodies was assessed with negative controls using pre-immune sera.
Secondary antibodies were Cy3-conjugated anti-sheep and Cy5-conjugated anti-rabbit (1:1000, Jackson Immunoresearch). After washing with PBS and briefly with water, the sections were mounted on glass slides with fluoromount g DAPI (Southern Biotech, Birmingham, AL, USA) and visualized using a Leica SP5 confocal microscope (Leica, Wetzlar, Germany).

Chen J., Lin X., Bhattacharya S., Wiesehöfer C., Wennemuth G., Müller K, & Montag D. (2023). Neuroplastin Expression in Male Mice Is Essential for Fertility, Mating, and Adult Testosterone Levels. International Journal of Molecular Sciences, 25(1), 177.

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