Category Archives: Follicles - Part 2

Distribution of the «1 to «6 Chains of Type IV Collagen: RESULTS(2)

Type IV Collagen a1 and a2 in Antral Follicles Antibodies to the a1 and a2 chains of type IV collagen in antral follicles produced similar staining patterns (Fig. 2). In all of the follicles examined for a1(IV) staining and all … Continue reading

Distribution of the «1 to «6 Chains of Type IV Collagen: RESULTS(1)

There are many structures in the ovary that contain basal laminae, but comments in this paper will be confined to ovarian follicles and their basal laminae. Specific staining for each of the individual type IV collagen chains was observed in … Continue reading

Distribution of the «1 to «6 Chains of Type IV Collagen: MATERIALS AND METHODS(8)

Samples were denatured by the addition of an equal volume of 0.05 M Tris buffer, pH 6.8, containing 2% SDS, 10% glycerol, 2 mM EDTA, 0.005% bromophenol blue, 0.1 M 4,4’diaminodiphenyl sulfone, 12 M urea solution, at 100°C for 5 … Continue reading

Distribution of the «1 to «6 Chains of Type IV Collagen: MATERIALS AND METHODS(7)

Images were captured using the AX70 microscope via a video linkage to an Apple Macintosh (Cupertino, CA) computer utilizing the program NIH Image 1.60b7 or were photographed using the Vanox microscope with Olympus C35AD-4 camera attachment. Photographs used in all … Continue reading

Distribution of the «1 to «6 Chains of Type IV Collagen: MATERIALS AND METHODS(6)

These were conjugated either with different fluo-rophores or with one conjugated to biotin-SP to allow further amplification by incubation with streptavidin conjugated to either Cy3 or FITC. Control sections for dual staining were as follows: the relevant polyclonal primary antibody … Continue reading

Distribution of the «1 to «6 Chains of Type IV Collagen: MATERIALS AND METHODS(5)

Sections were then washed (four times, 5 min each in hPBS) and incubated with the appropriate fluorophore-con-jugated secondary antibodies for 2 h. To achieve further amplification of the immunostaining, in some instances sections were instead incubated with secondary antisera conjugated … Continue reading

Distribution of the «1 to «6 Chains of Type IV Collagen: MATERIALS AND METHODS(4)

Fluorophore-conjugated secondary antibodies used in this study were from Jackson ImmunoResearch Laboratories (West Grove, PA) and included Cy3-conjugated AffiniPure donkey anti-rat IgG (cat. #712-165-153), Cy3-conjugated AffiniPure donkey anti-mouse IgG (cat. #715165-150), and fluorescein (DTAF)-conjugated AffiniPure donkey anti-rabbit IgG (cat. #711-095-152), … Continue reading

Distribution of the «1 to «6 Chains of Type IV Collagen: MATERIALS AND METHODS(3)

The a5 antibody was raised against the collagenase-resistant residue of human glomerulus basement membrane. It was screened by ELISA and SDS-PAGE to recombinant human type IV collagen chains and further characterized by indirect immunofluorescence within human kidney sections. This detects … Continue reading

Distribution of the «1 to «6 Chains of Type IV Collagen: MATERIALS AND METHODS(2)

Antibodies The current study used both rat monoclonal antibodies and mouse monoclonal antibodies directed against individual a chains of type IV collagen. The rat monoclonal antibodies were to each of the type IV collagen a1 to a6 chains, and they … Continue reading

Distribution of the «1 to «6 Chains of Type IV Collagen: MATERIALS AND METHODS(1)

Tissues All bovine tissue was collected at a local abattoir, within 20 min of slaughter, from cows assessed visually as being nonpregnant. A slice of up to 5 mm was cut through the center of ovaries (n = 10) to … Continue reading

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