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- Validations
- Western blot [2]
- Immunocytochemistry [2]
- Immunohistochemistry [8]
- Flow cytometry [1]
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- Product number
- PA5-114387 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SEMA3B Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 500 µg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SEMA3B in the following samples: Lane 1: rat thymus tissues lysates, Lane 2: rat kidney tissue lysates, Lane 3: mouse thymus tissue lysates, Lane 4: mouse lung tissue lysates, Lane 5: mouse small intestine tissue lysates, Lane 6: mouse kidney tissue lysates, Lane 7: mouse HEPA1-6 whole cell lysates. Samples consisting of 50 µg (reducing conditions) of protein was separated with 5-20% SDS-PAGE gel (70V, Stacking gel; 90V, Resolving gel; 2-3 hrs.), transferred to a Nitrocellulose membrane (150mA, 50-90 min) and washed with TBS-0.1% Tween (3 times, 5 minutes/wash) and blocked with 5% Non-fat Milk/TBS (1.5 hrs., room temperature). The membrane was incubated in SEMA3B polyclonal antibody (Product # PA5-114387) at a dilution of 0.5 µg/mL (overnight, 4°C), followed by goat anti-rabbit IgG-HRP and chemiluminescence (ECL) with a dilution of 1:10,000 (1.5 hours, room temperature).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SEMA3B in the following samples: Lane 1: human placenta tissue lysates, Lane 2: human U2OS whole cell lysates, Lane 3: human A549 whole cell lysates, Lane 4: human U-87MG whole cell lysates. Samples consisting of 50 µg (reducing conditions) of protein was separated with 5-20% SDS-PAGE gel (70V, Stacking gel; 90V, Resolving gel; 2-3 hrs.), transferred to a Nitrocellulose membrane (150mA, 50-90 min) and washed with TBS-0.1% Tween (3 times, 5 minutes/wash) and blocked with 5% Non-fat Milk/TBS (1.5 hrs., room temperature). The membrane was incubated in SEMA3B polyclonal antibody (Product # PA5-114387) at a dilution of 0.5 µg/mL (overnight, 4°C), followed by goat anti-rabbit IgG-HRP and chemiluminescence (ECL) with a dilution of 1:10,000 (1.5 hours, room temperature).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry analysis of SEMA3B in A431 cells. Antigen retrieval was performed with enzyme antigen retrieval (15 min). Samples were blocked with 10% goat serum and incubated in SEMA3B polyclonal antibody (Product # PA5-114387) at a dilution of 2 µg/mL (overnight, 4°C), followed by DyLight 488 conjugated goat anti-rabbit IgG (30 min, 37°C) and DAPI at a dilution of 1:100.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry analysis of SEMA3B in A431 cells. Antigen retrieval was performed with enzyme antigen retrieval (15 min). Samples were blocked with 10% goat serum and incubated in SEMA3B polyclonal antibody (Product # PA5-114387) at a dilution of 2 µg/mL (overnight, 4°C), followed by DyLight 488 conjugated goat anti-rabbit IgG (30 min, 37°C) and DAPI at a dilution of 1:100.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SEMA3B in paraffin-embedded mouse brain tissue. Antigen retrieval was performed with EDTA buffer (pH 8.0, epitope retrieval solution). Samples were blocked with 10% goat serum and incubated in SEMA3B polyclonal antibody (Product # PA5-114387) at a dilution of 1 µg/mL (overnight, 4°C), followed by biotinylated goat anti-rabbit IgG (30 min, 37°C) and Strepavidin-Biotin-Complex (SABC) with DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SEMA3B in paraffin-embedded human mammary cancer tissue. Antigen retrieval was performed with EDTA buffer (pH 8.0, epitope retrieval solution). Samples were blocked with 10% goat serum and incubated in SEMA3B polyclonal antibody (Product # PA5-114387) at a dilution of 1 µg/mL (overnight, 4°C), followed by biotinylated goat anti-rabbit IgG (30 min, 37°C) and Strepavidin-Biotin-Complex (SABC) with DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SEMA3B in paraffin-embedded rat brain tissue. Antigen retrieval was performed with EDTA buffer (pH 8.0, epitope retrieval solution). Samples were blocked with 10% goat serum and incubated in SEMA3B polyclonal antibody (Product # PA5-114387) at a dilution of 1 µg/mL (overnight, 4°C), followed by biotinylated goat anti-rabbit IgG (30 min, 37°C) and Strepavidin-Biotin-Complex (SABC) with DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SEMA3B in paraffin-embedded rat brain tissue. Antigen retrieval was performed with EDTA buffer (pH 8.0, epitope retrieval solution). Samples were blocked with 10% goat serum and incubated in SEMA3B polyclonal antibody (Product # PA5-114387) at a dilution of 1 µg/mL (overnight, 4°C), followed by biotinylated goat anti-rabbit IgG (30 min, 37°C) and Strepavidin-Biotin-Complex (SABC) with DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SEMA3B in paraffin-embedded mouse brain tissue. Antigen retrieval was performed with EDTA buffer (pH 8.0, epitope retrieval solution). Samples were blocked with 10% goat serum and incubated in SEMA3B polyclonal antibody (Product # PA5-114387) at a dilution of 1 µg/mL (overnight, 4°C), followed by biotinylated goat anti-rabbit IgG (30 min, 37°C) and Strepavidin-Biotin-Complex (SABC) with DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SEMA3B in paraffin-embedded mouse brain tissue. Antigen retrieval was performed with EDTA buffer (pH 8.0, epitope retrieval solution). Samples were blocked with 10% goat serum and incubated in SEMA3B polyclonal antibody (Product # PA5-114387) at a dilution of 1 µg/mL (overnight, 4°C), followed by biotinylated goat anti-rabbit IgG (30 min, 37°C) and Strepavidin-Biotin-Complex (SABC) with DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SEMA3B in paraffin-embedded human mammary cancer tissue. Antigen retrieval was performed with EDTA buffer (pH 8.0, epitope retrieval solution). Samples were blocked with 10% goat serum and incubated in SEMA3B polyclonal antibody (Product # PA5-114387) at a dilution of 1 µg/mL (overnight, 4°C), followed by biotinylated goat anti-rabbit IgG (30 min, 37°C) and Strepavidin-Biotin-Complex (SABC) with DAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of SEMA3B in paraffin-embedded human colon cancer tissue. Antigen retrieval was performed with EDTA buffer (pH 8.0, epitope retrieval solution). Samples were blocked with 10% goat serum and incubated in SEMA3B polyclonal antibody (Product # PA5-114387) at a dilution of 1 µg/mL (overnight, 4°C), followed by biotinylated goat anti-rabbit IgG (30 min, 37°C) and Strepavidin-Biotin-Complex (SABC) with DAB.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of SEMA3B in A549 cells. Samples were blocked with 10% normal goat serum and incubated in SEMA3B polyclonal antibody (Product # PA5-114387) at a dilution of 1 µg/1x10^6 cells (30 min, 20°C), followed by DyLight 488 conjugated goat anti-rabbit IgG (30 min, 20°C) with a dilution of 5-10 µg/1x10^6 cells (30 min, 20°C). Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) and unlabeled sample (Red line).
