EBs in fresh EB medium were pre-incubated with rabbit anti-Laminin 1+2 (abcam) or rat anti-Perlecan (Millipore) at 10 µg/ml for 2 hrs at 37°C in 7.5% CO2. The EBs were then harvested, washed in PBS, fixed with 4% PFA and sectioned as described in the ‘Immunofluorescence analysis’ protocol. 0.2% Triton X-100 permeabilized sections were subsequently treated with appropriate fluorophore-conjugated secondary antibodies to visualize any pre-incubated antibodies present in the sections.
Rabbit anti laminin 1 2
Manufactured by Abcam
Rabbit anti-Laminin 1+2 is a primary antibody that recognizes the Laminin 1 and Laminin 2 proteins. Laminin is a major structural component of the basement membrane. This antibody can be used to detect and localize Laminin 1 and Laminin 2 in various sample types.
Automatically generated - may contain errors
Lab products found in correlation
Rat anti perlecan, by Merck Group (1 mentions)
Anti cd3 alexafluor 647, by BioLegend (1 mentions)
Anti b220 brilliant violet 421, by BioLegend (1 mentions)
Lsm 710 axioobserver, by Zeiss (1 mentions)
Volocity v6, by PerkinElmer (1 mentions)
Prolong diamond antifade, by Thermo Fisher Scientific (1 mentions)
Zen 2012, by Zeiss (1 mentions)
2 protocols using rabbit anti laminin 1 2
1
Preincubation of EBs for Immunofluorescence
2
Immunofluorescent Tissue Staining and Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Spleens and lymph nodes were fixed for 2–4 h in 4% PFA in PBS, washed, and incubated overnight in 30% sucrose in PBS. Tissues were embedded in OCT and sectioned using a cryostat. Sections were blocked for 30 min in staining solution (5% donkey serum in PBS-Tween) containing FcRblock and subsequently incubated overnight at 4°C in staining solution with 1–2 ng/µl anti–CD3-AlexaFluor 647 (Clone 17A2, Biolegend; Cat. #100209), anti-B220-Brilliant Violet 421 (Clone RA3-6B2, Biolegend; Cat. #103239), and rabbit anti-laminin 1+2 (Abcam; Cat. #AB7463) labeled with CF568 Mix-n-stain kit (Biotium). After washing in PBS, slides were mounted using ProLong Diamond antifade (Invitrogen) and imaged at room temperature using a Zeiss LSM710 AxioObserver inverted confocal microscope. Images were captured using Zen 2012 (Zeiss) software and analyzed with Volocity v6.3 (Perkin Elmer) software with γ = 1.00.
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