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prm2 14500 1 ap proteintech  (Proteintech)


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    Structured Review

    Proteintech prm2 14500 1 ap proteintech
    Prm2 14500 1 Ap Proteintech, supplied by Proteintech, used in various techniques. Bioz Stars score: 91/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+prm2/PRM2+Antibody/pm41866391-112-45-47
    Average 91 stars, based on 15 article reviews
    prm2 14500 1 ap proteintech - by Bioz Stars, 2026-10
    91/100 stars

    Images

    Related Articles

    Immunostaining:

    Article Title: Panoramic transcriptome analysis and functional screening of long noncoding RNAs in mouse spermatogenesis
    Article Snippet: Finally, the slides were mounted with SlowFadeTM Gold Antifade Mountant with DAPI (Invitrogen) and observed under LSM 780 confocal microscope (Zeiss) for fluorescent signal analysis. .. Primary antibodies used in immunostaining were as follows: anti-ZBTB16 (Santa cruz, sc-28319), anti-HOOK1 (santa cruz, sc- 398233), anti-KIT (Cell Signalling Technology, 3074), Anti- H2AX (Abcam, ab2893), anti-SYCP3 (Abcam, ab97672), anti-WT1 (Abcam, ab89901), anti-CLGN (Abcam, ab172477), Alexa FluorTM 594 Conjugated-Lectin PNA (Invitrogen, L32459), anti- TNP1 (Proteintech, 17178-1-AP), anti-PRM2 (Proteintech, 14500-1-AP). .. Intensity and number of cells with staining were calculated using image analysis software of ZEISS ZEN lite 3.0; percentage of cells with staining per tubule was the ration of positive cell number/ nucleus number.

    Immunofluorescence:

    Article Title: A 22-amino-acid peptide regulates tight junctions through occludin and cell apoptosis.
    Article Snippet: .. Types and sources of antibodies Origin Primary antibody: anti-Bax (rat origin) Abcam, USA Primary antibody: anti-WT1 (rabbit origin) Abcam, USA Primary antibody: anti-Prm2 (rabbit origin) Proteintech, USA Secondary antibodies: anti-Alexa Fluor CY3 (anti-mouse) Abcam, USA Secondary antibody: anti-Alexa Fluor 488 (anti-rabbit) Abcam, USA Immunofluorescence primary antibody (diluted) Beyotime, China Immunofluorescence secondary antibody (diluted) Beyotime, China PBS ZSGB-BIO, Chia Paraformaldehyde Shanghai Sangong Goat serum HyClone, USA DAPI staining solution Beyotime, China Anti-fluorescence quenching agent Beyotime, China Primary antibody: occludin (rabbit origin) Abcam, USA Secondary antibody: Anti-Alexa Fluor cy3 (anti-rabbit) Abcam, USA Immunofluorescence primary antibody (diluted) Beyotime, China Immunofluorescence secondary antibody (diluted) Beyotime, China PBS ZSGB-BIO, Chia Paraformaldehyde Shanghai Sangong Goat serum HyClone, USA DAPI staining solution Beyotime, China Anti-fluorescence quenching agent Beyotime, China dropwise, followed by incubation at 37 ◦C for 60 min in a humidified box. ..

    Article Title: H3K36me2 methyltransferase NSD2 orchestrates epigenetic reprogramming during spermatogenesis
    Article Snippet: .. Antibodies used for western blot (WB), immunofluorescence (IF) and chromatin immunoprecipitation (ChIP) were as follows: anti-H3K36me3 (Solarbio no. K003337P, 1:400 for IF), anti-H3K36me2 (Solarbio no. K003336P, 1:400 for IF), anti-H3 (Solarbio no. K107283P, 1:800 for WB), anti-γ-H2A.X (Solarbio no. K006207P, 1:200 for IF, 1:1000 for WB), anti-mCherry (Solarbio no. K200015M, 1:1000 for WB), anti-H4K16ac (Abcam no. ab109463, 1:200 for IF), anti-NSD2 (Abcam no. ab-75359, 1:1000 for WB, 1:200 for IF), anti-PSMA8 (Proteintech no. 4022-1-AP, 1:1000 for WB), anti-GAPDH (Proteintech no. 60004-1-Ig, 1:1000 for WB), anti-PRM2 (Briar Patch Biosciences, Hup2B, 1:200 for IF, 1:800 for WB), anti-WT1 (Abcam no. ab89901, 1:200 for IF), anti-H4K5ac (ABclonal no. A19525, 1:1000 for WB, 1:200 for IF), anti-H4K8ac (ABclonal no. A7258, 1:1000 for WB, 1:200 for IF), anti-EP300 (Abcam no. ab-10485, 1:1000 for WB), anti-H4 (ABclonal no. A1131, 1:1000 for WB) and anti-α-Tubulin (Proteintech no. 66031-1-Ig, 1:1000 for WB). .. Mouse testes and epididymis were collected and fixed in Bouin’s solution (Sigma, HT10132) at 4°C overnight and then washed with 75% alcohol five times, 30 min each time.

    Staining:

    Article Title: A 22-amino-acid peptide regulates tight junctions through occludin and cell apoptosis.
    Article Snippet: .. Types and sources of antibodies Origin Primary antibody: anti-Bax (rat origin) Abcam, USA Primary antibody: anti-WT1 (rabbit origin) Abcam, USA Primary antibody: anti-Prm2 (rabbit origin) Proteintech, USA Secondary antibodies: anti-Alexa Fluor CY3 (anti-mouse) Abcam, USA Secondary antibody: anti-Alexa Fluor 488 (anti-rabbit) Abcam, USA Immunofluorescence primary antibody (diluted) Beyotime, China Immunofluorescence secondary antibody (diluted) Beyotime, China PBS ZSGB-BIO, Chia Paraformaldehyde Shanghai Sangong Goat serum HyClone, USA DAPI staining solution Beyotime, China Anti-fluorescence quenching agent Beyotime, China Primary antibody: occludin (rabbit origin) Abcam, USA Secondary antibody: Anti-Alexa Fluor cy3 (anti-rabbit) Abcam, USA Immunofluorescence primary antibody (diluted) Beyotime, China Immunofluorescence secondary antibody (diluted) Beyotime, China PBS ZSGB-BIO, Chia Paraformaldehyde Shanghai Sangong Goat serum HyClone, USA DAPI staining solution Beyotime, China Anti-fluorescence quenching agent Beyotime, China dropwise, followed by incubation at 37 ◦C for 60 min in a humidified box. ..

    Article Title: Anatomical Transcriptome Atlas of the Male Mouse Reproductive System During Aging
    Article Snippet: After washed with PBS then incubated with HRP Conjugated Goat Anti-Rabbit IgG (1:100, CW0103, CWBIO, Beijing, China) at room temperature for 30 min, the sections were counterstained with Hematoxylin Staining Solution (ZLI-9610, ZSGB-BIO, Beijing, China). .. For immunofluorescent staining in mouse epididymis and human testis, the primary antibody included anti-Pla2g2d (1:100, PA5-93147, Invitrogen, United States) and anti-PRM2 (1:100, 14500-1-AP, Proteintech Group, Inc, China) were used. .. After washed with PBS then incubated with CY3 Conjugated AffiniPure Goat Anti-rabbit IgG (1:100, BA1032, Boster Biological Technology co.ltd, Wuhan, China) at room temperature for 30 min, the sections were counterstained with DAPI (10 μg/ml, C1002, Beyotime Biotechnology, Shanghai, China).

    Article Title: Anatomical Transcriptome Atlas of the Male Mouse Reproductive System During Aging.
    Article Snippet: After washed with PBS then incubated with HRP Conjugated Goat Anti-Rabbit IgG (1: 100, CW0103, CWBIO, Beijing, China) at room temperature for 30 min, the sections were counterstained with Hematoxylin Staining Solution (ZLI-9610, ZSGB-BIO, Beijing, China). .. For immunofluorescent staining in mouse epididymis and human testis, the primary antibody included anti-Pla2g2d (1:100, PA593147, Invitrogen, United States) and anti-PRM2 (1:100, 14500- 1-AP, Proteintech Group, Inc, China) were used. .. After washed with PBS then incubated with CY3 Conjugated AffiniPure Goat Anti-rabbit IgG (1:100, BA1032, Boster Biological Technology co.ltd, Wuhan, China) at room temperature for 30 min, the sections were counterstained with DAPI (10 μg/ml, C1002, Beyotime Biotechnology, Shanghai, China).

    Incubation:

    Article Title: A 22-amino-acid peptide regulates tight junctions through occludin and cell apoptosis.
    Article Snippet: .. Types and sources of antibodies Origin Primary antibody: anti-Bax (rat origin) Abcam, USA Primary antibody: anti-WT1 (rabbit origin) Abcam, USA Primary antibody: anti-Prm2 (rabbit origin) Proteintech, USA Secondary antibodies: anti-Alexa Fluor CY3 (anti-mouse) Abcam, USA Secondary antibody: anti-Alexa Fluor 488 (anti-rabbit) Abcam, USA Immunofluorescence primary antibody (diluted) Beyotime, China Immunofluorescence secondary antibody (diluted) Beyotime, China PBS ZSGB-BIO, Chia Paraformaldehyde Shanghai Sangong Goat serum HyClone, USA DAPI staining solution Beyotime, China Anti-fluorescence quenching agent Beyotime, China Primary antibody: occludin (rabbit origin) Abcam, USA Secondary antibody: Anti-Alexa Fluor cy3 (anti-rabbit) Abcam, USA Immunofluorescence primary antibody (diluted) Beyotime, China Immunofluorescence secondary antibody (diluted) Beyotime, China PBS ZSGB-BIO, Chia Paraformaldehyde Shanghai Sangong Goat serum HyClone, USA DAPI staining solution Beyotime, China Anti-fluorescence quenching agent Beyotime, China dropwise, followed by incubation at 37 ◦C for 60 min in a humidified box. ..

    Western Blot:

    Article Title: H3K36me2 methyltransferase NSD2 orchestrates epigenetic reprogramming during spermatogenesis
    Article Snippet: .. Antibodies used for western blot (WB), immunofluorescence (IF) and chromatin immunoprecipitation (ChIP) were as follows: anti-H3K36me3 (Solarbio no. K003337P, 1:400 for IF), anti-H3K36me2 (Solarbio no. K003336P, 1:400 for IF), anti-H3 (Solarbio no. K107283P, 1:800 for WB), anti-γ-H2A.X (Solarbio no. K006207P, 1:200 for IF, 1:1000 for WB), anti-mCherry (Solarbio no. K200015M, 1:1000 for WB), anti-H4K16ac (Abcam no. ab109463, 1:200 for IF), anti-NSD2 (Abcam no. ab-75359, 1:1000 for WB, 1:200 for IF), anti-PSMA8 (Proteintech no. 4022-1-AP, 1:1000 for WB), anti-GAPDH (Proteintech no. 60004-1-Ig, 1:1000 for WB), anti-PRM2 (Briar Patch Biosciences, Hup2B, 1:200 for IF, 1:800 for WB), anti-WT1 (Abcam no. ab89901, 1:200 for IF), anti-H4K5ac (ABclonal no. A19525, 1:1000 for WB, 1:200 for IF), anti-H4K8ac (ABclonal no. A7258, 1:1000 for WB, 1:200 for IF), anti-EP300 (Abcam no. ab-10485, 1:1000 for WB), anti-H4 (ABclonal no. A1131, 1:1000 for WB) and anti-α-Tubulin (Proteintech no. 66031-1-Ig, 1:1000 for WB). .. Mouse testes and epididymis were collected and fixed in Bouin’s solution (Sigma, HT10132) at 4°C overnight and then washed with 75% alcohol five times, 30 min each time.

    Chromatin Immunoprecipitation:

    Article Title: H3K36me2 methyltransferase NSD2 orchestrates epigenetic reprogramming during spermatogenesis
    Article Snippet: .. Antibodies used for western blot (WB), immunofluorescence (IF) and chromatin immunoprecipitation (ChIP) were as follows: anti-H3K36me3 (Solarbio no. K003337P, 1:400 for IF), anti-H3K36me2 (Solarbio no. K003336P, 1:400 for IF), anti-H3 (Solarbio no. K107283P, 1:800 for WB), anti-γ-H2A.X (Solarbio no. K006207P, 1:200 for IF, 1:1000 for WB), anti-mCherry (Solarbio no. K200015M, 1:1000 for WB), anti-H4K16ac (Abcam no. ab109463, 1:200 for IF), anti-NSD2 (Abcam no. ab-75359, 1:1000 for WB, 1:200 for IF), anti-PSMA8 (Proteintech no. 4022-1-AP, 1:1000 for WB), anti-GAPDH (Proteintech no. 60004-1-Ig, 1:1000 for WB), anti-PRM2 (Briar Patch Biosciences, Hup2B, 1:200 for IF, 1:800 for WB), anti-WT1 (Abcam no. ab89901, 1:200 for IF), anti-H4K5ac (ABclonal no. A19525, 1:1000 for WB, 1:200 for IF), anti-H4K8ac (ABclonal no. A7258, 1:1000 for WB, 1:200 for IF), anti-EP300 (Abcam no. ab-10485, 1:1000 for WB), anti-H4 (ABclonal no. A1131, 1:1000 for WB) and anti-α-Tubulin (Proteintech no. 66031-1-Ig, 1:1000 for WB). .. Mouse testes and epididymis were collected and fixed in Bouin’s solution (Sigma, HT10132) at 4°C overnight and then washed with 75% alcohol five times, 30 min each time.



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    Image Search Results


    (A) Schematic depiction of murine PRM1 and PRM2 amino acid sequences (grey box = cP2). (B) Schematic depiction of CRISPR/Cas9-mediated gene editing of the Prm1-Prm2 locus. Black arrow heads indicate the target sites of CRISPR-Cas9 guide RNAs. (C) Schematic depiction of Prm1 and Prm2 gene-edited alleles and their respective amino acid predictions. (D) Genomic WT and dHET DNA amplified with PCRs targeting either Prm1 or Prm2 . L = ladder. (E) Average pregnancy frequency after mating WT and dHET males with WT C57BL/6J females. Dots represent different males (n = number of males). (F) Average litter size after mating WT and dHET males with WT females. Dots show each litter size (n = number of males).

    Journal: bioRxiv

    Article Title: Male mice heterozygous for Protamine-1 and Protamine-2 are infertile displaying sperm damage and retention of Protamine-2 precursors, transition proteins and histones

    doi: 10.64898/2026.03.15.711850

    Figure Lengend Snippet: (A) Schematic depiction of murine PRM1 and PRM2 amino acid sequences (grey box = cP2). (B) Schematic depiction of CRISPR/Cas9-mediated gene editing of the Prm1-Prm2 locus. Black arrow heads indicate the target sites of CRISPR-Cas9 guide RNAs. (C) Schematic depiction of Prm1 and Prm2 gene-edited alleles and their respective amino acid predictions. (D) Genomic WT and dHET DNA amplified with PCRs targeting either Prm1 or Prm2 . L = ladder. (E) Average pregnancy frequency after mating WT and dHET males with WT C57BL/6J females. Dots represent different males (n = number of males). (F) Average litter size after mating WT and dHET males with WT females. Dots show each litter size (n = number of males).

    Article Snippet: Primary antibodies diluted in blocking solution were added to the membrane and incubated overnight at 4 °C [PRM1 (Briar Patch Biosciences; Mab-Hup1N-150; 1:1000), PRM2 (Briar Patch Biosciences; Mab-Hup2B-150; 1:1000), ODF2 (proteintech; 12058-1-AP; 1:500-1:1000), histone H3 (abcam; ab1791; 1:1000), histone H4 (abcam; ab177840 ; 1:500)].

    Techniques: CRISPR, Amplification

    (A) Separation of basic cauda epididymal sperm protein extractions on Coomassie-stained acid-urea (AU) and thiourea gels. PRMs are detected at the bottom of the gel. Precursor bands of PRM2 are prominent in dHET samples (dashed box). Thiourea gels were used for analyses shown in (B-I). (B) Relative level of protamines compared to total protein content extracted from WT and dHET sperm. (C) Relative level of total PRM2 compared to total protein content extracted from WT and dHET sperm. (D) Relative level of PRM1 compared to total protein content extracted from WT and dHET sperm. (E) Relative level of mP2 compared to total protein content extracted from WT and dHET sperm. (F) Relative level of PRM2 precursors compared to total protein content extracted from WT and dHET sperm. (G) Relative level of mP2 compared to total PRM content extracted from WT and dHET sperm. (H) Relative level of PRM2 precursors compared to total PRM content extracted from WT and dHET sperm. (I) Relative level of total PRM2 precursors compared to total PRM content extracted from WT and dHET sperm. (J) Relative amount of PRM2 precursors comared to total PRM2 extracted from WT and dHET sperm. (K) Separation of basic protein extractions from whole testis on thiourea gels stained with Coomassie. PRM2 precursors are marked with a dashed box. Thiourea gels were used for quantifications shown in (L-S). (L) Relative level of protamines compared to total protein content extracted from WT and dHET testis. (M) Relative level of total PRM2 compared to total protein content extracted from WT and dHET testis. (N) Relative level of PRM1 compared to total protein content extracted from WT and dHET testis. (O) Relative level of mP2 compared to total protein content extracted from WT and dHET testis. (P) Relative level of PRM2 precursors compared to total protein content extracted from WT and dHET testis. (Q) Relative level of mP2 compared to total PRM content extracted from WT and dHET testis. (R) Relative level of PRM2 precursors compared to total PRM content extracted from WT and dHET testis. (S) Relative level of total PRM2 precursors compared to total PRM content extracted from WT and dHET testis. (T) Relative amount of PRM2 precursors comared to total PRM2 extracted from WT and dHET testis.

    Journal: bioRxiv

    Article Title: Male mice heterozygous for Protamine-1 and Protamine-2 are infertile displaying sperm damage and retention of Protamine-2 precursors, transition proteins and histones

    doi: 10.64898/2026.03.15.711850

    Figure Lengend Snippet: (A) Separation of basic cauda epididymal sperm protein extractions on Coomassie-stained acid-urea (AU) and thiourea gels. PRMs are detected at the bottom of the gel. Precursor bands of PRM2 are prominent in dHET samples (dashed box). Thiourea gels were used for analyses shown in (B-I). (B) Relative level of protamines compared to total protein content extracted from WT and dHET sperm. (C) Relative level of total PRM2 compared to total protein content extracted from WT and dHET sperm. (D) Relative level of PRM1 compared to total protein content extracted from WT and dHET sperm. (E) Relative level of mP2 compared to total protein content extracted from WT and dHET sperm. (F) Relative level of PRM2 precursors compared to total protein content extracted from WT and dHET sperm. (G) Relative level of mP2 compared to total PRM content extracted from WT and dHET sperm. (H) Relative level of PRM2 precursors compared to total PRM content extracted from WT and dHET sperm. (I) Relative level of total PRM2 precursors compared to total PRM content extracted from WT and dHET sperm. (J) Relative amount of PRM2 precursors comared to total PRM2 extracted from WT and dHET sperm. (K) Separation of basic protein extractions from whole testis on thiourea gels stained with Coomassie. PRM2 precursors are marked with a dashed box. Thiourea gels were used for quantifications shown in (L-S). (L) Relative level of protamines compared to total protein content extracted from WT and dHET testis. (M) Relative level of total PRM2 compared to total protein content extracted from WT and dHET testis. (N) Relative level of PRM1 compared to total protein content extracted from WT and dHET testis. (O) Relative level of mP2 compared to total protein content extracted from WT and dHET testis. (P) Relative level of PRM2 precursors compared to total protein content extracted from WT and dHET testis. (Q) Relative level of mP2 compared to total PRM content extracted from WT and dHET testis. (R) Relative level of PRM2 precursors compared to total PRM content extracted from WT and dHET testis. (S) Relative level of total PRM2 precursors compared to total PRM content extracted from WT and dHET testis. (T) Relative amount of PRM2 precursors comared to total PRM2 extracted from WT and dHET testis.

    Article Snippet: Primary antibodies diluted in blocking solution were added to the membrane and incubated overnight at 4 °C [PRM1 (Briar Patch Biosciences; Mab-Hup1N-150; 1:1000), PRM2 (Briar Patch Biosciences; Mab-Hup2B-150; 1:1000), ODF2 (proteintech; 12058-1-AP; 1:500-1:1000), histone H3 (abcam; ab1791; 1:1000), histone H4 (abcam; ab177840 ; 1:500)].

    Techniques: Staining

    (A) Testis to body weight ratio of dHET and WT mice (n= number of males). (B) Representative stage VII-VIII seminiferous tubule section of dHET and WT stained with Hematoxylin and Eosin. (C) Percentages of viable and inviable mature sperm of WT, Prm1 +/- , Prm1 -/- , Prm2 +/- , Prm2 -/- and dHET mice. Part of the data has been published before , . Data are mean ± s.d. and was analyzed by ANOVA. (D) Percentages of motile and immotile sperm in WT and dHET mature swim-out samples (n= number of males). (E) Quantification of axonemal integrity. A minimum of 100 flagella cross sections were per animal were analyzed (n= number of males). Data are mean ± s.d. and were analyzed by two-tailed, unpaired Student’s t-test (***p<0.001). (F-N) Sperm stained with DAPI (blue), PNA-FITC (green) and MitoTracker (red). Turquoise arrows highlight excess residual cytoplasm. (C). Scale bars: 50 μm (B); 10 μm (F-N).

    Journal: bioRxiv

    Article Title: Male mice heterozygous for Protamine-1 and Protamine-2 are infertile displaying sperm damage and retention of Protamine-2 precursors, transition proteins and histones

    doi: 10.64898/2026.03.15.711850

    Figure Lengend Snippet: (A) Testis to body weight ratio of dHET and WT mice (n= number of males). (B) Representative stage VII-VIII seminiferous tubule section of dHET and WT stained with Hematoxylin and Eosin. (C) Percentages of viable and inviable mature sperm of WT, Prm1 +/- , Prm1 -/- , Prm2 +/- , Prm2 -/- and dHET mice. Part of the data has been published before , . Data are mean ± s.d. and was analyzed by ANOVA. (D) Percentages of motile and immotile sperm in WT and dHET mature swim-out samples (n= number of males). (E) Quantification of axonemal integrity. A minimum of 100 flagella cross sections were per animal were analyzed (n= number of males). Data are mean ± s.d. and were analyzed by two-tailed, unpaired Student’s t-test (***p<0.001). (F-N) Sperm stained with DAPI (blue), PNA-FITC (green) and MitoTracker (red). Turquoise arrows highlight excess residual cytoplasm. (C). Scale bars: 50 μm (B); 10 μm (F-N).

    Article Snippet: Primary antibodies diluted in blocking solution were added to the membrane and incubated overnight at 4 °C [PRM1 (Briar Patch Biosciences; Mab-Hup1N-150; 1:1000), PRM2 (Briar Patch Biosciences; Mab-Hup2B-150; 1:1000), ODF2 (proteintech; 12058-1-AP; 1:500-1:1000), histone H3 (abcam; ab1791; 1:1000), histone H4 (abcam; ab177840 ; 1:500)].

    Techniques: Staining, Two Tailed Test

    (A) Representative images of DAPI-stained WT and dHET mature sperm. Sperm are visualized twice with different exposure times (8 ms and 33 ms), showing examples of DAPI-bright and DAPI-weak sperm in a dHET sample. Scale bars: 50 μm. (B) Sperm nuclei consensus shapes of WT and dHET epididymal sperm. Only DAPI-bright dHET cells were included (n = 3 males per genotype). (C-E) Violin plots depicting the area (C), minimum diameter (D) and length of hook (E) of WT and dHET epididymal sperm. (F) Agarose gel loaded with genomic DNA isolated from WT, Prm1 +/- (P1 +/- ), Prm1 -/- (P1 -/- ), Prm2 +/- (P2 +/- ), Prm2 -/- (P2 -/- ), and dHET cauda epididymal sperm. Part of the data has been published before . (G) IHC against 8-OHdG on epididymal (caput and cauda) tissue from WT and dHET males (Dapi counterstain in grey) Scale bars: 50 μm. (H) Percentage of 8-OHdG-positive sperm from WT and dHET sperm from caput and cauda tissue (n = 3 males per genotype). (I-K) Representative TEM images from (I) WT and (J-K) dHET mature sperm. Scale bars: 1 μm. (J) Ratio of slightly damaged (as shown in J) and heavily damaged (as shown in K) dHET sperm. Data are mean ± s.d. and were analyzed by two-tailed, unpaired Student’s t-test (***p<0.001). (M) Quantification of the difference in grey scale (grey scale range) measured for epididymal sperm nuclei from WT and dHET males (n = number of males).

    Journal: bioRxiv

    Article Title: Male mice heterozygous for Protamine-1 and Protamine-2 are infertile displaying sperm damage and retention of Protamine-2 precursors, transition proteins and histones

    doi: 10.64898/2026.03.15.711850

    Figure Lengend Snippet: (A) Representative images of DAPI-stained WT and dHET mature sperm. Sperm are visualized twice with different exposure times (8 ms and 33 ms), showing examples of DAPI-bright and DAPI-weak sperm in a dHET sample. Scale bars: 50 μm. (B) Sperm nuclei consensus shapes of WT and dHET epididymal sperm. Only DAPI-bright dHET cells were included (n = 3 males per genotype). (C-E) Violin plots depicting the area (C), minimum diameter (D) and length of hook (E) of WT and dHET epididymal sperm. (F) Agarose gel loaded with genomic DNA isolated from WT, Prm1 +/- (P1 +/- ), Prm1 -/- (P1 -/- ), Prm2 +/- (P2 +/- ), Prm2 -/- (P2 -/- ), and dHET cauda epididymal sperm. Part of the data has been published before . (G) IHC against 8-OHdG on epididymal (caput and cauda) tissue from WT and dHET males (Dapi counterstain in grey) Scale bars: 50 μm. (H) Percentage of 8-OHdG-positive sperm from WT and dHET sperm from caput and cauda tissue (n = 3 males per genotype). (I-K) Representative TEM images from (I) WT and (J-K) dHET mature sperm. Scale bars: 1 μm. (J) Ratio of slightly damaged (as shown in J) and heavily damaged (as shown in K) dHET sperm. Data are mean ± s.d. and were analyzed by two-tailed, unpaired Student’s t-test (***p<0.001). (M) Quantification of the difference in grey scale (grey scale range) measured for epididymal sperm nuclei from WT and dHET males (n = number of males).

    Article Snippet: Primary antibodies diluted in blocking solution were added to the membrane and incubated overnight at 4 °C [PRM1 (Briar Patch Biosciences; Mab-Hup1N-150; 1:1000), PRM2 (Briar Patch Biosciences; Mab-Hup2B-150; 1:1000), ODF2 (proteintech; 12058-1-AP; 1:500-1:1000), histone H3 (abcam; ab1791; 1:1000), histone H4 (abcam; ab177840 ; 1:500)].

    Techniques: Staining, Agarose Gel Electrophoresis, Isolation, Two Tailed Test

    (A) CMA3 staining of mature WT and dHET sperm counterstained by DAPI. Scale bars: 30 μm. (B) Quantification of CMA3-stained nuclei in WT and dHET samples. Data are mean ± s.d. and were analyzed by two-tailed, unpaired Student’s t-test (***p<0.001). (C) Representative image of a Coomassie stained AU-PAGE of basic nuclear-enriched proteins isolated from WT and dHET epididymal sperm. Bands corresponding to ODF2 and protamines are marked. Bands containing predominantly PRM1 and mature PRM2 (mP2) are labelled and areas of bands containing Prm2 precursors are marked (red dashed box). Non-protamine, nuclear-enriched proteins are found to be differentially abundant in dHET compared to WT sperm (red box). (D) Upper part of image shown in (C) compared to merged Western Blots against ODF2, histone H3 and histone H4 (pan-H3 and pan-H4 antibodies). (E) Western blots against histone H3 and H4 basic protein extractions of WT testis lysate and sperm samples form WT, dHET, Prm1 +/- (P1 +/- ), Prm1 -/- (P1 -/- ), Prm2 +/- (P2 +/- ), Prm2 -/- (P2 -/- ). ODF2 was used as loading control, as described before . (F) IHC against pre-PRM2 (antibody epitope in cP2) on WT and dHET epididymal caput tissue sections. (G) IHC against transition proteins TNP1 and TNP2 on WT and dHET epididymal caput tissue sections. Scales (F-G): 50 µm

    Journal: bioRxiv

    Article Title: Male mice heterozygous for Protamine-1 and Protamine-2 are infertile displaying sperm damage and retention of Protamine-2 precursors, transition proteins and histones

    doi: 10.64898/2026.03.15.711850

    Figure Lengend Snippet: (A) CMA3 staining of mature WT and dHET sperm counterstained by DAPI. Scale bars: 30 μm. (B) Quantification of CMA3-stained nuclei in WT and dHET samples. Data are mean ± s.d. and were analyzed by two-tailed, unpaired Student’s t-test (***p<0.001). (C) Representative image of a Coomassie stained AU-PAGE of basic nuclear-enriched proteins isolated from WT and dHET epididymal sperm. Bands corresponding to ODF2 and protamines are marked. Bands containing predominantly PRM1 and mature PRM2 (mP2) are labelled and areas of bands containing Prm2 precursors are marked (red dashed box). Non-protamine, nuclear-enriched proteins are found to be differentially abundant in dHET compared to WT sperm (red box). (D) Upper part of image shown in (C) compared to merged Western Blots against ODF2, histone H3 and histone H4 (pan-H3 and pan-H4 antibodies). (E) Western blots against histone H3 and H4 basic protein extractions of WT testis lysate and sperm samples form WT, dHET, Prm1 +/- (P1 +/- ), Prm1 -/- (P1 -/- ), Prm2 +/- (P2 +/- ), Prm2 -/- (P2 -/- ). ODF2 was used as loading control, as described before . (F) IHC against pre-PRM2 (antibody epitope in cP2) on WT and dHET epididymal caput tissue sections. (G) IHC against transition proteins TNP1 and TNP2 on WT and dHET epididymal caput tissue sections. Scales (F-G): 50 µm

    Article Snippet: Primary antibodies diluted in blocking solution were added to the membrane and incubated overnight at 4 °C [PRM1 (Briar Patch Biosciences; Mab-Hup1N-150; 1:1000), PRM2 (Briar Patch Biosciences; Mab-Hup2B-150; 1:1000), ODF2 (proteintech; 12058-1-AP; 1:500-1:1000), histone H3 (abcam; ab1791; 1:1000), histone H4 (abcam; ab177840 ; 1:500)].

    Techniques: Staining, Two Tailed Test, Isolation, Western Blot, Control

    A The Kmal in total proteins of (TP) and protein fractions of the mitochondrion (Mit), head, and cytoplasm (Cyt) isolated from human sperm were examined by Western blot. The specificity of cellular components was validated by Western blot using the corresponding markers: PRM2 for sperm head, COX6B1 for mitochondrion, and ACTIN for cytoplasm. The proteins for Western blot were shown in the coomassie brilliant blue (CBB) staining image. B , C The localization of malonylated proteins within human sperm was investigated by immunofluorescence assay via laser scanning confocal microscopy (LSCM, ( B )) and super-resolution structured illumination microscopy (SIM, ( C )). All the experiments were conducted with samples from 8 normozoospermic men. The scale bar represents 5 μm.

    Journal: Communications Biology

    Article Title: Lysine malonylation regulates human sperm motility

    doi: 10.1038/s42003-026-09683-y

    Figure Lengend Snippet: A The Kmal in total proteins of (TP) and protein fractions of the mitochondrion (Mit), head, and cytoplasm (Cyt) isolated from human sperm were examined by Western blot. The specificity of cellular components was validated by Western blot using the corresponding markers: PRM2 for sperm head, COX6B1 for mitochondrion, and ACTIN for cytoplasm. The proteins for Western blot were shown in the coomassie brilliant blue (CBB) staining image. B , C The localization of malonylated proteins within human sperm was investigated by immunofluorescence assay via laser scanning confocal microscopy (LSCM, ( B )) and super-resolution structured illumination microscopy (SIM, ( C )). All the experiments were conducted with samples from 8 normozoospermic men. The scale bar represents 5 μm.

    Article Snippet: Anti-ACTIN antibody (66009-1-Ig), anti-SIRT5 antibody (67257-1-Ig), anti-acetyl-CoA carboxylase 1 (ACC1) antibody (21923-1-AP), anti-fatty acid synthase (FASN) antibody (10624-2-AP), anti-protamine 2 (PRM2) antibody (14500-1-AP), anti-GAPDHS (83290-3-RR) and anti-VDAC3 (82666-14-RR) were obtained from Proteintech Group, Inc. (Rosemont, IL, USA).

    Techniques: Isolation, Western Blot, Staining, Immunofluorescence, Confocal Microscopy, Microscopy