mouse monoclonal anti anxa5  (Boster Bio)

Journal: International journal of biochemistry & physiology

Article Title: Fibrinogen-Related Protein, Fgl2, of Hamster Cauda Epididymal Fluid: Enzymatic Characterization, and Identification of Fgl2-Binding Proteins and Ligand of Defective Hamster Sperm Organelles

doi: 10.23880/ijbp-16000228

Figure Lengend Snippet: Immunoblot of Annexin V (I): Western blots of cauda epididymal fluid (lane 1) and death cocoon fraction (lane 2) fractionated by SDS-PAGE stained with anti-Annexin V monoclonal antibody. Annexin V immunoreactive band was detected by LICOR Odyssey CVX imaging system. Each lane contains 5μg of protein. Immunolocalization of Annexin V (II): Immunofluorescence (Panel A) and a matched phase contrast (Panel A’) photomicrographs of intact hamster cauda epididymal spermatozoa immunostained with anti-Annexin V monoclonal antibody. Note that the tails of two defective spermatozoa (as shown by the arrow) are stained with an annexin V antibody (Panel A).

Article Snippet: Immunoblots were blocked with TBS (0.15 M NaCl, 20 mM Tris-HCl Buffer, pH 7.5) containing 0.1% Tween 20 and 1% BSA and then incubated with immune serum (rabbit polyclonal anti-FGL2/ rabbit monoclonal fibrinogen beta chain [abcam] /mouse monoclonal Annexin V/ ANXA5 [abcam] antibodies) or nonimmune serum diluted in TBS containing 0.1% Tween 20 (TBS-TW) and 1% BSA.

Techniques: Western Blot, SDS Page, Staining, Imaging, Immunofluorescence

Journal: International journal of biochemistry & physiology

Article Title: Fibrinogen-Related Protein, Fgl2, of Hamster Cauda Epididymal Fluid: Enzymatic Characterization, and Identification of Fgl2-Binding Proteins and Ligand of Defective Hamster Sperm Organelles

doi: 10.23880/ijbp-16000228

Figure Lengend Snippet: Triton X-100 soluble fraction of cauda sperm (50μg protein) was immunoprecipitated with anti-FGL2 and fractionated by SDS-PAGE on a 12% gel followed by immunoblot analyses. Immunoreactive bands were detected by LICOR Odyssey CVX imaging system. Sperm lysate lanes reveal the presence of FGL2 (A-lane 1) and Annexin V (B-lane 3). As shown in figure A-lane 2, FGL2 was recovered in the anti-FGL2 immunoprecipitated (IP) pellet, whereas Annexin V co-precipitated with FGL2 (Figure B-lane 4).

Article Snippet: Immunoblots were blocked with TBS (0.15 M NaCl, 20 mM Tris-HCl Buffer, pH 7.5) containing 0.1% Tween 20 and 1% BSA and then incubated with immune serum (rabbit polyclonal anti-FGL2/ rabbit monoclonal fibrinogen beta chain [abcam] /mouse monoclonal Annexin V/ ANXA5 [abcam] antibodies) or nonimmune serum diluted in TBS containing 0.1% Tween 20 (TBS-TW) and 1% BSA.

Techniques: Immunoprecipitation, SDS Page, Western Blot, Imaging

Journal: International journal of biochemistry & physiology

Article Title: Fibrinogen-Related Protein, Fgl2, of Hamster Cauda Epididymal Fluid: Enzymatic Characterization, and Identification of Fgl2-Binding Proteins and Ligand of Defective Hamster Sperm Organelles

doi: 10.23880/ijbp-16000228

Figure Lengend Snippet: Panel A shows the localization of Annexin V in the flagellum of nitrogen- cavitated cauda sperm; no staining was observed in heads. A matched phase contrast micrograph (Panel A’) reveals the presence of detached heads and tails. Panel B reveals the presence of FGL2 in detached heads and tails of nitrogen-cavitated cauda sperm when immunostained with anti-FGL2; no staining was evident in intact spermatozoa (as shown by arrow) in a companion phase contrast micrograph (Panel B’). Panel C shows a fluorescence micrograph of isolated tails stained with anti-Annexin V. An intense staining was evident in the flagellum, and no fluorescence was observed on heads (Panel C). Few sperm heads are present in isolated tails, as shown in the companion phase contrast micrograph (Panel C’). Bar = 50 μm.

Article Snippet: Immunoblots were blocked with TBS (0.15 M NaCl, 20 mM Tris-HCl Buffer, pH 7.5) containing 0.1% Tween 20 and 1% BSA and then incubated with immune serum (rabbit polyclonal anti-FGL2/ rabbit monoclonal fibrinogen beta chain [abcam] /mouse monoclonal Annexin V/ ANXA5 [abcam] antibodies) or nonimmune serum diluted in TBS containing 0.1% Tween 20 (TBS-TW) and 1% BSA.

Techniques: Staining, Fluorescence, Isolation