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Ab57577

Manufactured by Abcam
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Ab57577 is a primary antibody product manufactured by Abcam. It is a recombinant rabbit monoclonal antibody targeted against a specific protein epitope. The antibody is supplied in a buffered solution.

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Lab products found in correlation

Lsm 710, by Zeiss (3 mentions) Ab77256, by Abcam (3 mentions) Oct4a, by Cell Signaling Technology (1 mentions) Lin28a, by Cell Signaling Technology (1 mentions) Ventana benchmark xt system, by Roche (1 mentions) M4403, by Merck Group (1 mentions) Ab97959, by Abcam (1 mentions) M7315, by Agilent Technologies (1 mentions) Ki67 sp6, by Cell Marque (1 mentions) M7240, by Agilent Technologies (1 mentions) Ab128874, by Abcam (1 mentions) A2547, by Merck Group (1 mentions) Sc 5565, by Santa Cruz Biotechnology (1 mentions) Mab4337, by Merck Group (1 mentions)

7 protocols using ab57577

1

Histological Analysis of Ovarian Cell Colonies

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Histology of the colonies was performed after mechanically detaching of whole MEF layer including the ovarian cell colonies from the cell culture well. After detachment, the randomly arranged MEF layer was fixed in Bouin's solution for 3 hours, washed several times for at least 24 h in 70% EtOH, and then embedded in paraffin wax. The resulting cell culture conglomerate was randomly sectioned. Due to the large number of colonies sufficient sections of ovarian cell colonies in different orientations were available for histological analysis. Immunohistochemistry was performed as described recently [18 (link)] using the following primary antibodies: OCT4A (#2890S, Cell Signaling Technology, Germany; 1 : 100), LIN28A (#3978S, Cell Signaling Technology; 1 : 100–1 : 200), SALL4 (#ab57577, Abcam, UK; 1 : 200), and VASA (DDX4; #AF2030, R&D Systems, Germany; 1 : 100).
Fereydouni B., Salinas-Riester G., Heistermann M., Dressel R., Lewerich L., Drummer C, & Behr R. (2015). Long-Term Oocyte-Like Cell Development in Cultures Derived from Neonatal Marmoset Monkey Ovary. Stem Cells International, 2016, 2480298.
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2

Tissue Immunohistochemistry Protocol

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Tissue samples were fixed in 4% buffered formaldehyde, dehydrated, embedded in paraffin, and sectioned at 2 µm. H&E staining was performed according to standard laboratory protocols. Immunohistochemical stainings were performed on a Ventana BenchMark XT system (Roche Diagnostics). The following primary antibodies were used: SALL4 (ab57577, abcam, 1:50), Ki67 (SP6, Cell Marque, 1:750), SOX2 (ab97959, abcam, 1:1000), MAP2C (M4403, Sigma–Aldrich, 1:3000), CD56 (MSK006, Zytomed, 1:2000), Synaptophysin (M7315, Dako, 1:500). As a chromogen, 3,3ʹ-diaminobenzine (DAB) was used.
Dottermusch M., Biabani A., Lempertz T., Schumann Y., Navolic J., Godbole S., Obrecht D., Frank S., Dorostkar M.M., Voß H., Schlüter H., Rutkowski S., Schüller U, & Neumann J.E. (2023). Integrated proteomics spotlight the proteasome as a therapeutic vulnerability in embryonal tumors with multilayered rosettes. Neuro-Oncology, 26(5), 935-949.
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3

SALL4 Knockdown and Immunofluorescence Imaging

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40 h after SALL4 KD with shRNA 1 and 2, SNU398 liver cancer cells were fixed with 4% paraformaldehyde in PBS for 15 min. Fixed cells were permeabilized with 0.1% Triton-x in PBS, washed with PBS containing 0.1% Tween-20, and blocked with 3% bovine serum albumin (BSA). Primary antibodies against HP1 (Abcam ab77256) or SALL4 (Abcam ab57577) were incubated with cells overnight at 4 degrees, in PBS containing 0.3% BSA and 0.1% Tween-20. The next day, cells were washed with Tween-20/PBS, incubated with secondary anti-goat or anti-rabbit antibodies conjugated to Alexa Fluorophore 594 at room temperature for an hour. Cells were washed and stained with DAPI DNA stain for 5 min and mounted with Vectashield mounting medium. Images were taken with a confocal microscope (Zeiss LSM710) with the same settings for all samples.
Kong N.R., Bassal M.A., Tan H.K., Kurland J.V., Yong K.J., Young J.J., Yang Y., Li F., Lee J.D., Liu Y., Wu C.S., Stein A., Luo H.R., Silberstein L.E., Bulyk M.L., Tenen D.G, & Chai L. (2021). Zinc Finger Protein SALL4 Functions through an AT-Rich Motif to Regulate Gene Expression. Cell reports, 34(1), 108574.
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4

SALL4 Knockdown and Immunofluorescence Imaging

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40 h after SALL4 KD with shRNA 1 and 2, SNU398 liver cancer cells were fixed with 4% paraformaldehyde in PBS for 15 min. Fixed cells were permeabilized with 0.1% Triton-x in PBS, washed with PBS containing 0.1% Tween-20, and blocked with 3% bovine serum albumin (BSA). Primary antibodies against HP1 (Abcam ab77256) or SALL4 (Abcam ab57577) were incubated with cells overnight at 4 degrees, in PBS containing 0.3% BSA and 0.1% Tween-20. The next day, cells were washed with Tween-20/PBS, incubated with secondary anti-goat or anti-rabbit antibodies conjugated to Alexa Fluorophore 594 at room temperature for an hour. Cells were washed and stained with DAPI DNA stain for 5 min and mounted with Vectashield mounting medium. Images were taken with a confocal microscope (Zeiss LSM710) with the same settings for all samples.
Kong N.R., Bassal M.A., Tan H.K., Kurland J.V., Yong K.J., Young J.J., Yang Y., Li F., Lee J.D., Liu Y., Wu C.S., Stein A., Luo H.R., Silberstein L.E., Bulyk M.L., Tenen D.G, & Chai L. (2021). Zinc Finger Protein SALL4 Functions through an AT-Rich Motif to Regulate Gene Expression. Cell reports, 34(1), 108574.
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5

Immunohistochemical Profiling of Hepatoblastoma

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Tissue samples were fixed in 10% neutral buffered formalin and embedded in paraffin. The specimens were cut at a thickness of 3 μm from paraffin‐embedded conventional blocks and deparaffinized with xylene and ethanol. Hepatoblastoma foci were stained to detect the expression of Ki‐67 (mouse monoclonal; M7240, DAKO), Iba1 (rabbit polyclonal; 019‐19741, WAKO), CD163 (mouse monoclonal; AM3K, Transgenic, Kumamoto, Japan), SALL4 (mouse monoclonal; Abcam, Cambridge, UK, ab57577), bromodomain‐containing protein 4 (Brd4, rabbit monoclonal; Abcam, ab128874), and interleukin (IL)‐34 (mouse monoclonal; 1D12, Abcam). Horseradish peroxidase‐labeled goat anti‐rabbit or anti‐mouse antibodies (Nichirei) were used as secondary antibodies.
Irie T., Yoshii D., Komohara Y., Fujiwara Y., Kadohisa M., Honda M., Suzu S., Matsuura T., Kohashi K., Oda Y, & Hibi T. (2022). IL‐34 in hepatoblastoma cells potentially promote tumor progression via autocrine and paracrine mechanisms. Cancer Medicine, 11(6), 1441-1453.
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6

SALL4 Knockdown Immunofluorescence Imaging

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40 hours after SALL4 KD with shRNA 1 and 2, SNU398 liver cancer cells were fixed with 4% paraformaldehyde in PBS for 15 minutes. Fixed cells were permeabilized with 0.1% Triton-x in PBS, washed with PBS containing 0.1% Tween-20, and blocked with 3% bovine serum albumin (BSA). Primary antibodies against HP1 (Abcam ab77256) or SALL4 (Abcam ab57577) were incubated with cells overnight at 4 degrees, in PBS containing 0.3% BSA and 0.1% Tween-20. The next day, cells were washed with Tween-20/PBS, incubated with secondary anti-goat or anti-rabbit antibodies conjugated to Alexa Fluorophore 594 at room temperature for an hour. Cells were washed and stained with DAPI DNA stain for 5 minutes and mounted with Vectashield mounting medium. Images were taken with a confocal microscope (Zeiss LSM710) with the same settings for all samples.
Kong N.R., Bassal M.A., Tan H.K., Kurland J.V., Yong K.J., Young J.J., Yang Y., Li F., Lee J., Liu Y., Wu C., Stein A., Luo H.R., Silberstein L.E., Bulyk M.L., Tenen D.G., & Chai L. (2020). Zinc finger protein SALL4 functions through an AT-rich motif to regulate gene expression.
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7

Immunohistochemical Analysis of Testicular Tissue

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Processing of the testicular tissue and the staining protocol were as described elsewhere (26) . Primary antibodies used in this study included undifferentiated embryonic cell transcription factor I (UTF1, MAB4337, Millipore; dilution, 1:50), sal-like protein 4 (SALL4, ab57577, Abcam; dilution, 1:500), melanoma antigen family A4 (MAGE A4, provided by Professor G. C. Spagnoli from the University Hospital of Basel, Switzerland; dilution, 1:20), VIM (sc-5565, Santa Cruz Biotechnology; dilution, 1:50), and SMA (A2547, Sigma-Aldrich; dilution, 1:1,000). Incubation with corresponding immunoglobulin G (IgG) antibodies served as negative controls. The following day, sections were incubated with the corresponding horseradish peroxidase-conjugated secondary antibody at a dilution of 1:100, and staining was visualized by performing diaminobenzidine (Sigma-Aldrich) treatment followed by counterstaining with hematoxylin. As controls, testicular tissues from prostate cancer patients, who were orchiectomized for therapeutic reasons and showed normal spermatogenesis, were used.
Schneider F., Redmann K., Wistuba J., Schlatt S., Kliesch S, & Neuhaus N. (2015). Comparison of enzymatic digestion and mechanical dissociation of human testicular tissues. Fertility and sterility, 104(2).
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