Mouse brain was fixed in 4% formaldehyde for 24 h, and then incubated in 30% sucrose until tissues are sink. Fixed brain was flash frozen using pre-cooled isopentane (− 78 °C), sectioned at 30 μm using Microm HM525 Cryostat (Thermo) and picked up on Superfrost Plus slides (VWR, 48311-703). Sections were blocked with 5% normal goat serum and washed in PBS with 1% bovine serum albumin (BSA) and incubated with rabbit- anti-mouse Iba-1 primary antibody (FUJIFILM Wako Pure Chemical Corporation 019-19741, 1:500) or rabbit- anti-mouse Iba-1 primary antibody Alexa Fluor® 594 conjugate (Cell Signaling, 48934, 1:50) overnight. Sections were washed with phosphate buffer with 1% Tween 20 (PBS-T), and then incubated in goat anti-rabbit IgG (H + L) secondary antibody (Vector laboratory CY-1300, 1:250) at room temperature for 1 h when unconjugated antibody was used. Afterword, sections were washed three times with PBS-T followed by mounting on coverslip using Vectashield DAPI (4′6-diamidino-2-phenylindole 2HCl, Vector Labs, Burlingame, U.S.) mounting media to detect nuclei.
Goat anti rabbit igg h l secondary antibody
Manufactured by Vector Laboratories
Sourced in United States
Goat anti-rabbit IgG (H+L) secondary antibody is a reagent used in immunoassays and other immunological techniques. It is designed to detect and bind to rabbit primary antibodies, providing a means to amplify and visualize the signal from the primary antibody-antigen interaction.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 594 conjugate, by Cell Signaling Technology (2 mentions)
Microm hm 525 cryostat, by Thermo Fisher Scientific (2 mentions)
Rabbit anti mouse iba 1 primary antibody, by Fujifilm (2 mentions)
Vectashield dapi 4 6 diamidino 2 phenylindole 2hcl, by Vector Laboratories (2 mentions)
Superfrost plus slides, by Avantor (2 mentions)
Normal goat serum (ngs), by Vector Laboratories (1 mentions)
Triton x 100, by Thermo Fisher Scientific (1 mentions)
3 3 diaminobenzidine (dab), by Agilent Technologies (1 mentions)
Haematoxylin, by Merck Group (1 mentions)
Vectastain elite abc kit, by Vector Laboratories (1 mentions)
3 protocols using goat anti rabbit igg h l secondary antibody
1
Immunohistochemical Analysis of Microglia in Mouse Brain
2
Immunohistochemical Analysis of Survivin Expression
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As previously described (Yamada et al, 2010 (link)), paraffin-embedded prostate tissue sections were deparaffinised and rehydrated and then subjected to antigen retrieval in 10 mM sodium citrate buffer at 95°C. Slides were then incubated in 1% H2O2 in methanol to block the endogenous peroxidase activity. Sections were blocked with 10% normal goat serum (Vector Laboratories, Burlingame, CA, USA) and 0.3% Triton X-100 (Fisher Scientific, Pittsburgh, PA, USA) in TBS for 1 h at room temperature and incubated in survivin antibody overnight at 4°C. The next day, affinity-purified, biotinylated goat anti-rabbit IgG (H+L) secondary antibody (Vector Laboratories) was added to the sections and the staining was visualised with detection reagents: VECTASTAIN Elite ABC kit (Vector Laboratories) and DAB (DAKO, Carpinteria, CA, USA). The slides were counterstained with haematoxylin (Sigma-Aldrich, St Louis, MO, USA), rinsed and dehydrated before cover slips were placed over the tissue.
3
Immunohistochemical Analysis of Microglial Activation
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Mouse brain was fixed in 4% formaldehyde for 24 hours, and then incubated in 30% sucrose until tissues are sink. Fixed brain was flash frozen using pre-cooled isopentane (−78 °C), sectioned at 30 μm using Microm HM525 Cryostat (Thermo) and picked up on Superfrost Plus slides (VWR, 48311–703). Sections were blocked with 5% normal goat serum and washed in PBS with 1% bovine serum albumin (BSA) and incubated with rabbit- anti-mouse Iba-1 primary antibody (FUJIFILM Wako Pure Chemical Corporation 019–19741, 1:500) or rabbit- anti-mouse Iba-1 primary antibody Alexa Fluor® 594 conjugate (Cell Signaling, 48934, 1:50) overnight. Sections were washed with phosphate buffer with 1% Tween 20 (PBS-T), and then incubated in goat anti-rabbit IgG (H+L) secondary antibody (Vector laboratory CY-1300, 1:250) at room temperature for 1 hour when unconjugated antibody was used. Afterword, sections were washed three times with PBS-T followed by mounting on coverslip using Vectashield DAPI (4′6-diamidino-2-phenylindole 2HCl, Vector Labs, Burlingame, U.S.) mounting media to detect nuclei.
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