hCG was from NV Organon (Oss, Holland). Human recombinant insulin (Humulin NPH) was from Eli Lilly (Lilly Pharmaceuticals, Giza, Egypt). 3,3-diaminobenzidine tetrahydrochloride (DAB) was from Sigma-Aldrich (St. Louis, MO). The avidin-biotinylated-peroxidase complex detection system (ABC kit) was from Vector Laboratories Inc. (Burlingame, CA). The primary antibodies used for Western blot and immunohistochemical analyses in the present study, their dilution, and sources are listed in Supplemental Table 1 . Anti-mouse IgG horseradish peroxidase (HRP)-conjugated goat (A2304), and anti-rabbit IgG HRP-conjugated goat (A0545) secondary antibodies were from Sigma-Aldrich. Alexa Fluor 594-conjugated goat polyclonal anti-mouse IgG and Alexa Fluor 488-conjugated goat polyclonal anti-rabbit IgG were from Invitrogen (UK).
Alexa fluor 488 conjugated goat polyclonal anti rabbit igg
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
Alexa Fluor 488-conjugated goat polyclonal anti-rabbit IgG is a secondary antibody that binds to rabbit immunoglobulin G (IgG) and is conjugated to the Alexa Fluor 488 fluorescent dye. This product is designed for use in various immunodetection techniques, such as Western blotting, immunohistochemistry, and flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Anti mouse igg horseradish peroxidase hrp conjugated goat a2304, by Merck Group (2 mentions)
Anti rabbit igg hrp conjugated goat a0545, by Merck Group (2 mentions)
Alexa fluor 594 conjugated goat polyclonal anti mouse igg, by Thermo Fisher Scientific (2 mentions)
Avidin biotinylated peroxidase complex detection system abc kit, by Vector Laboratories (2 mentions)
3 3 diaminobenzidine tetrahydrochloride dab, by Merck Group (1 mentions)
Human chorionic gonadotropin (hcg), by Organon (1 mentions)
Humulin nph, by Eli Lilly (1 mentions)
Diaminobenzidine dab, by Merck Group (1 mentions)
Ar sc 816, by Santa Cruz Biotechnology (1 mentions)
Flutamide, by Merck Group (1 mentions)
P21cip1, by Santa Cruz Biotechnology (1 mentions)
Apoptag fluorescein direct in situ apoptosis detection kit, by Merck Group (1 mentions)
Ab66155, by Abcam (1 mentions)
Bz 9000 all in one fluorescence microscope, by Keyence (1 mentions)
Vectashield mounting medium, by Vector Laboratories (1 mentions)
Mouse monoclonal anti actin antibody, by Abcam (1 mentions)
4 protocols using alexa fluor 488 conjugated goat polyclonal anti rabbit igg
1
Immunohistochemical and Western Blot Analyses
2
Molecular Mechanisms of Androgen Receptor Regulation
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DHT (a specific AR agonist, D-5027), flutamide (a specific AR antagonist, F-0397), and diaminobenzidine (DAB) were from Sigma-Aldrich (St. Louis, MO). Avidin-biotinylated-peroxidase complex detection system (ABC kit) was from Vector Laboratories Inc. (Burlingame, CA). All antibodies were from the following sources: AR (#5153, this antibody reacts with human tissues), AMPKα (#2532), phospho-AMPKα (phosphorylated threonine 172, #2535), and Ki-67 (#11882) antibodies were from Cell Signaling Technology (Danvers, MA); AR (sc-816, this antibody reacts with rodent tissues) and p21Cip1 antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA); and cysteine-rich protein 61 (CYR61, HPA029853) antibody was from Sigma-Aldrich. Anti-mouse IgG horseradish peroxidase (HRP)-conjugated goat (A2304) and anti-rabbit IgG HRP-conjugated goat (A0545) secondary antibodies were from Sigma-Aldrich. Alexa Fluor 594-conjugated goat polyclonal anti-mouse IgG, and Alexa Fluor 488-conjugated goat polyclonal anti-rabbit IgG were from Invitrogen (Life Technologies Europe BV, Stockholm, Sweden).
3
Histological Analysis of XTC Tumors
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XTC.UC1 and XTC.UC1/MIEAP tumors were excised and fixed in 10% neutral‐buffered formalin and embedded in paraffin. Staining with H&E was carried out as mentioned above. For Ki‐67 staining, 4‐μm sections were treated with 3% hydrogen peroxide to block endogenous peroxidase and subjected to antigen retrieval by microwave treatment in citrate buffer, followed by incubation with rabbit polyclonal anti‐Ki‐67 Ab (ab66155, 1:200 dilution; Abcam) and then with Alexa Fluor 488‐conjugated goat polyclonal anti‐rabbit IgG (A‐11008, 1:200 dilution; Life Technologies). The slides were then analyzed with an All‐in‐one Fluorescence Microscope BZ‐9000 (Keyence). One hundred cells were evaluated to determine the percentage of Ki67‐positive cells (n = 4).
The TUNEL staining was carried out with the Apop‐tag Fluorescein Direct in situ apoptosis detection kit (Merck Millipore). The slides were embedded with VECTASHIELD Mounting Medium containing DAPI (Vector Laboratories) and analyzed using an All‐in‐One BZ‐9000 fluorescence microscope. One hundred cells were evaluated in each sample (n = 4) to determine the percentages of TUNEL‐positive cells.
The TUNEL staining was carried out with the Apop‐tag Fluorescein Direct in situ apoptosis detection kit (Merck Millipore). The slides were embedded with VECTASHIELD Mounting Medium containing DAPI (Vector Laboratories) and analyzed using an All‐in‐One BZ‐9000 fluorescence microscope. One hundred cells were evaluated in each sample (n = 4) to determine the percentages of TUNEL‐positive cells.
4
Immunohistochemical Staining and Western Blot Analysis of ABCC11 and Pepsinogen I
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For IHC staining, primary antibodies were used at the following concentrations: rabbit
polyclonal anti-human ABCC11/MRP8 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA), 1:200
and mouse monoclonal anti‑human pepsinogen I antibody (Sanbio BV, Uden, The Netherlands),
1:500. Horseradish peroxidase (HRP)-conjugated goat polyclonal anti-rabbit IgG (Nichirei
Biosciences, Tokyo, Japan) and HRP-conjugated goat poly‐clonal anti-mouse IgG (Nichirei
Biosciences) were used as detecting antibodies. Alexa Fluor 488-conjugated goat polyclonal
anti-rabbit IgG (Life Technologies), 1:2000 and Alexa Fluor 594-conjugated goat polyclonal anti-mouse IgG (Life Technologies), 1:2000 were also used as detecting antibodies in the IF assay. In the Western blot analysis, additional mouse monoclonal anti-actin antibody (Abcam, Cambridge, MA, USA) or anti-GAPDH antibody were used as internal controls.
polyclonal anti-human ABCC11/MRP8 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA), 1:200
and mouse monoclonal anti‑human pepsinogen I antibody (Sanbio BV, Uden, The Netherlands),
1:500. Horseradish peroxidase (HRP)-conjugated goat polyclonal anti-rabbit IgG (Nichirei
Biosciences, Tokyo, Japan) and HRP-conjugated goat poly‐clonal anti-mouse IgG (Nichirei
Biosciences) were used as detecting antibodies. Alexa Fluor 488-conjugated goat polyclonal
anti-rabbit IgG (Life Technologies), 1:2000 and Alexa Fluor 594-conjugated goat polyclonal anti-mouse IgG (Life Technologies), 1:2000 were also used as detecting antibodies in the IF assay. In the Western blot analysis, additional mouse monoclonal anti-actin antibody (Abcam, Cambridge, MA, USA) or anti-GAPDH antibody were used as internal controls.
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