biotin sp affinipure f ab 2 fragment donkey anti goat igg  (Jackson Immuno)

 
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    Name:
    Biotin SP AffiniPure F ab 2 Fragment Donkey Anti Goat IgG
    Description:
    F ab 2 fragment antibodies are generated by pepsin digestion of whole IgG antibodies to remove most of the Fc region while leaving some of the hinge region F ab 2 fragments have two antigen binding Fab portions linked together by disulfide bonds and therefore they are divalent The average molecular weight is about 110 kDa They are used for specific applications such as to avoid binding of secondary antibodies to live cells with Fc receptors or to Protein A or Protein G Based on immunoelectrophoresis and or ELISA the antibody reacts with whole molecule goat IgG It also reacts with the light chains of other goat immunoglobulins No antibody was detected against non immunoglobulin serum proteins The antibody has been tested by ELISA and or solid phase adsorbed to ensure minimal cross reaction with chicken guinea pig syrian hamster horse human mouse rabbit and rat serum proteins but it may cross react with immunoglobulins from other species
    Catalog Number:
    705-066-147
    Price:
    137
    Purity:
    The antibody was purified from antisera by a combination of pepsin digestion and immunoaffinity chromatography using antigens coupled to agarose beads. Fc fragments and whole IgG molecules have been removed.
    Conjugate:
    Biotin SP long spacer
    Size:
    ml
    Category:
    Secondary Antibody
    Source:
    Donkey
    Quantity:
    0 3
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    Structured Review

    Jackson Immuno biotin sp affinipure f ab 2 fragment donkey anti goat igg
    Biotin SP AffiniPure F ab 2 Fragment Donkey Anti Goat IgG
    F ab 2 fragment antibodies are generated by pepsin digestion of whole IgG antibodies to remove most of the Fc region while leaving some of the hinge region F ab 2 fragments have two antigen binding Fab portions linked together by disulfide bonds and therefore they are divalent The average molecular weight is about 110 kDa They are used for specific applications such as to avoid binding of secondary antibodies to live cells with Fc receptors or to Protein A or Protein G Based on immunoelectrophoresis and or ELISA the antibody reacts with whole molecule goat IgG It also reacts with the light chains of other goat immunoglobulins No antibody was detected against non immunoglobulin serum proteins The antibody has been tested by ELISA and or solid phase adsorbed to ensure minimal cross reaction with chicken guinea pig syrian hamster horse human mouse rabbit and rat serum proteins but it may cross react with immunoglobulins from other species
    https://www.bioz.com/result/biotin sp affinipure f ab 2 fragment donkey anti goat igg/product/Jackson Immuno
    Average 99 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    biotin sp affinipure f ab 2 fragment donkey anti goat igg - by Bioz Stars, 2020-04
    99/100 stars

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    Related Articles

    Transduction:

    Article Title: Endothelial CaMKII as a regulator of eNOS activity and NO-mediated vasoreactivity
    Article Snippet: Transduction Laboratories, anti-HA.11 antibody (clone 16B12, #MMS-101P) was from Covance. .. Goat anti-rabbit IgG (111-066-003) and donkey anti-goat IgG (705-066-147) biotinylated antibodies and normal rabbit and normal mouse IgG were from Jackson Immuno Research.

    Immunocytochemistry:

    Article Title: High Mobility Group 1 Protein (Hmg-1) Stimulates Proinflammatory Cytokine Synthesis in Human Monocytes
    Article Snippet: .. Secondary antibodies for immunocytochemistry included Fab2 -fragmented biotinylated donkey anti–goat IgG (cat. no. 705-066-147; Jackson ImmunoResearch Laboratories) diluted 1:1,000; biotinylated goat anti–mouse IgG1 (cat. no. M32115; Caltag Laboratories) diluted 1:600; or biotinylated goat anti–rat IgG (cat. no. BA9400; Vector Laboratories) diluted 1:500. .. Cytokine Detection by Indirect Immunocytochemistry.

    Article Title: Increased Feeding and Food Hoarding following Food Deprivation Are Associated with Activation of Dopamine and Orexin Neurons in Male Brandt's Voles
    Article Snippet: Another set of brain sections at 160 µm intervals was processed for Fos/orexin double immunocytochemistry. .. The same protocol outlined above was used except that the primary antibody for orexin labeling was goat anti-orexin (sc-8070; 1∶2000; Santa Cruz Biotechnology) and the secondary antibody was donkey anti-goat IgG (705-066-147; 1∶500; Jackson ImmunoResearch, West Grove, PA).

    Immunostaining:

    Article Title: Specific Connectivity and Unique Molecular Identity of MET Receptor Tyrosine Kinase-Expressing Serotonergic Neurons in the Caudal Dorsal Raphe Nuclei
    Article Snippet: Paragraph title: Immunostaining ... Secondary antibodies used were as follows: Alexa 647 AffiniPure F(ab′)2 Fragment Donkey Anti-Chicken IgG (Jackson ImmunoResearch Labs, Cat# 703-546-155), Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs, Cat# 705-066-147), DyLight 549 Streptavidin (Jackson ImmunoResearch Labs, Cat#016-580-084), Alexa 647 Donkey anti-Rabbit IgG (Thermo Fisher Scientific Cat# A-31573).

    Article Title: Distinct Projection Targets Define Subpopulations of Mouse Brainstem Vagal Neurons that Express the Autism-Associated MET Receptor Tyrosine Kinase
    Article Snippet: Paragraph title: Immunostaining ... After several rinses in PBS, tissue was then incubated for 2 hr at room temperature in PBS containing 2% normal donkey serum, 0.3% triton-X100, and one or more of the following species-specific secondary antibodies: Alexa Fluor® 488 AffiniPure F(ab′)2 Fragment Donkey Anti-Chicken IgG (Jackson ImmunoResearch Labs Cat# 703-546-155, RRID:AB_2340376), Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs Cat# 705-066-147, RRID:AB_2340398), DyLight 549 Streptavidin (Vector Laboratories Cat# SA-5549, RRID:AB_2336408), Alexa Fluor® 647 Donkey anti-Goat IgG (Thermo Fisher Scientific Cat# A-21447, RRID:AB_2535864), Cy3-AffiniPure Donkey Anti-Rabbit IgG, (Jackson ImmunoResearch Labs Cat# 711-165-152, RRID:AB_2307443), Alexa Fluor® 647 Donkey anti-Rabbit IgG, (Thermo Fisher Scientific Cat# A-31573, RRID:AB_2536183), and/or Alexa Fluor® 647 Donkey anti-Mouse IgG (Thermo Fisher Scientific Cat# A-31571, RRID:AB_2536181).

    Article Title: Distinct Projection Targets Define Subpopulations of Mouse Brainstem Vagal Neurons that Express the Autism-Associated MET Receptor Tyrosine Kinase
    Article Snippet: Sections were then washed several times in Blotto, and incubated for 1 hr at room temperature in Blotto containing Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Mouse IgG (Jackson ImmunoResearch Labs Cat# 715-066-150, RRID:AB_2340787) or Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs Cat# 705-066-147, RRID:AB_2340398). .. As a reference marker for neuronal subpopulations in the rhesus macaque brainstem, ChAT immunostaining was performed to localize cholinergic neurons in sections proximal to the GD151 brainstem sections used for MET immunostaining.

    Incubation:

    Article Title: Downstream Toll-like receptor signaling mediates adaptor-specific cytokine expression following focal cerebral ischemia
    Article Snippet: Secondary antibodies included biotin-conjugated donkey anti-rat (cat. no. 712-066-153) and donkey anti-goat (cat. no. 705-066-147) at 1:3,000 concentrations (both from Jackson Immunoresearch Laboratories, West Grove, PA, USA) for anti-Ly6B.2 and anti-CD45, respectively. .. Nonspecific immunoreactivity with primary antibodies was determined by the lack of DAB staining in negative controls incubated with normal immunoglobulin G derived from the species of the primary antibody.

    Article Title: Distinct Projection Targets Define Subpopulations of Mouse Brainstem Vagal Neurons that Express the Autism-Associated MET Receptor Tyrosine Kinase
    Article Snippet: .. After several rinses in PBS, tissue was then incubated for 2 hr at room temperature in PBS containing 2% normal donkey serum, 0.3% triton-X100, and one or more of the following species-specific secondary antibodies: Alexa Fluor® 488 AffiniPure F(ab′)2 Fragment Donkey Anti-Chicken IgG (Jackson ImmunoResearch Labs Cat# 703-546-155, RRID:AB_2340376), Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs Cat# 705-066-147, RRID:AB_2340398), DyLight 549 Streptavidin (Vector Laboratories Cat# SA-5549, RRID:AB_2336408), Alexa Fluor® 647 Donkey anti-Goat IgG (Thermo Fisher Scientific Cat# A-21447, RRID:AB_2535864), Cy3-AffiniPure Donkey Anti-Rabbit IgG, (Jackson ImmunoResearch Labs Cat# 711-165-152, RRID:AB_2307443), Alexa Fluor® 647 Donkey anti-Rabbit IgG, (Thermo Fisher Scientific Cat# A-31573, RRID:AB_2536183), and/or Alexa Fluor® 647 Donkey anti-Mouse IgG (Thermo Fisher Scientific Cat# A-31571, RRID:AB_2536181). .. After several rinses in PBS, sections were dried and a coverslip applied using fluoromount G mounting medium (Southern Biotech, Birmingham, AL).

    Article Title: TLR-9 and IL-15 Synergy Promotes the In Vitro Clonal Expansion of Chronic Lymphocytic Leukemia B Cells
    Article Snippet: The tissues were preblocked with 25% normal serum matching the secondary detection Ab’s species and further subjected to avidin/biotin blocking (kit from Vector Labs) before incubation at 4°C overnight in a humidified chamber with primary Ab: goat anti-human IL-15 (AF 315; R & D Systems), mouse anti-human Pax-5 (IS650, clone DAK; Dako), or mouse anti-human Ki-67 (N1633, clone Mib 1; Dako) at the recommended dilutions along with appropriate isotype controls. .. The following secondary biotin-conjugated, affinity-purified F(ab′)2 reagents were used for Ig detection (each commercially absorbed against human serum proteins): donkey anti-goat IgG (code 705-066-147; Jackson Immunoresearch), donkey anti-rabbit IgG (code 711-066-152; Jackson Immunoresearch), horse anti-mouse IgG (BA-2000; Vector Laboratories), or goat anti-mouse IgM (μ-chain–specific; code 115-066-075; Jackson Immunoresearch).

    Article Title: Distinct Projection Targets Define Subpopulations of Mouse Brainstem Vagal Neurons that Express the Autism-Associated MET Receptor Tyrosine Kinase
    Article Snippet: .. Sections were then washed several times in Blotto, and incubated for 1 hr at room temperature in Blotto containing Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Mouse IgG (Jackson ImmunoResearch Labs Cat# 715-066-150, RRID:AB_2340787) or Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs Cat# 705-066-147, RRID:AB_2340398). .. After washing in PBS, sections were incubated for 1 hr in Vectastain ABC Elite solution (Vector Labs, Burlingame, CA, RRID:AB_2336819), washed in PBS, and incubated for 4 minutes in ImmPACT DAB Peroxidase (HRP) Substrate (Vector Labs, Burlingame, CA, RRID:AB_2336520).

    Article Title: The Immune-Modulating Cytokine and Endogenous Alarmin Interleukin-33 Is Upregulated in Skin Exposed to Inflammatory UVB Radiation
    Article Snippet: .. After this, the TBS-washed slides were incubated for 1 hour at 37°C with a 1/200 biotin-conjugated donkey anti-goat IgG secondary antibody (705-066-147; Jackson Immunoresearch, West Grove, PA). .. The slides were then washed in TBS before a 30-minute incubation at 37°C in 1/1000 strepavidin Alexa Fluor 488 (S-11223; Molecular Probes, Carlsbad, CA).

    Article Title: Increased Feeding and Food Hoarding following Food Deprivation Are Associated with Activation of Dopamine and Orexin Neurons in Male Brandt's Voles
    Article Snippet: Sections were rinsed and incubated in avidin-biotin complex (Vector) in 0.1 M PBS for 90 min, and rinsed in 0.1 M PBS. .. The same protocol outlined above was used except that the primary antibody for orexin labeling was goat anti-orexin (sc-8070; 1∶2000; Santa Cruz Biotechnology) and the secondary antibody was donkey anti-goat IgG (705-066-147; 1∶500; Jackson ImmunoResearch, West Grove, PA).

    Article Title: YKL-40 Is Differentially Expressed in Human Embryonic Stem Cells and in Cell Progeny of the Three Germ Layers
    Article Snippet: .. Sections were then incubated with HNF-3β for 1½ hr at room temperature and then in biotin-SP-conjugated F(ab′)2 fragment donkey anti-goat (705-066-147; Jackson ImmunoResearch) followed by streptavidin-conjugated Dylight 594 (SA5594; Vector Laboratories) before being coverslipped. .. Prior to staining, nonspecific binding was inhibited by incubation for 30 min with 0.2% casein (C-7078; Sigma) at room temperature.

    Activity Assay:

    Article Title: Distinct Projection Targets Define Subpopulations of Mouse Brainstem Vagal Neurons that Express the Autism-Associated MET Receptor Tyrosine Kinase
    Article Snippet: Briefly, sections were washed several times in PBS and incubated for 5 min in 0.3% H2 O2 (Sigma, St. Louis, MO) in methanol to eliminate endogenous peroxidase activity. .. Sections were then washed several times in Blotto, and incubated for 1 hr at room temperature in Blotto containing Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Mouse IgG (Jackson ImmunoResearch Labs Cat# 715-066-150, RRID:AB_2340787) or Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs Cat# 705-066-147, RRID:AB_2340398).

    Western Blot:

    Article Title: Endothelial CaMKII as a regulator of eNOS activity and NO-mediated vasoreactivity
    Article Snippet: Anti-eNOS (# 610296) used for western blotting was from B.D. .. Goat anti-rabbit IgG (111-066-003) and donkey anti-goat IgG (705-066-147) biotinylated antibodies and normal rabbit and normal mouse IgG were from Jackson Immuno Research.

    Derivative Assay:

    Article Title: Downstream Toll-like receptor signaling mediates adaptor-specific cytokine expression following focal cerebral ischemia
    Article Snippet: Secondary antibodies included biotin-conjugated donkey anti-rat (cat. no. 712-066-153) and donkey anti-goat (cat. no. 705-066-147) at 1:3,000 concentrations (both from Jackson Immunoresearch Laboratories, West Grove, PA, USA) for anti-Ly6B.2 and anti-CD45, respectively. .. Nonspecific immunoreactivity with primary antibodies was determined by the lack of DAB staining in negative controls incubated with normal immunoglobulin G derived from the species of the primary antibody.

    Immunohistochemistry:

    Article Title: Downstream Toll-like receptor signaling mediates adaptor-specific cytokine expression following focal cerebral ischemia
    Article Snippet: Paragraph title: Immunohistochemistry ... Secondary antibodies included biotin-conjugated donkey anti-rat (cat. no. 712-066-153) and donkey anti-goat (cat. no. 705-066-147) at 1:3,000 concentrations (both from Jackson Immunoresearch Laboratories, West Grove, PA, USA) for anti-Ly6B.2 and anti-CD45, respectively.

    Article Title: TLR-9 and IL-15 Synergy Promotes the In Vitro Clonal Expansion of Chronic Lymphocytic Leukemia B Cells
    Article Snippet: Paragraph title: Immunohistochemical analyses ... The following secondary biotin-conjugated, affinity-purified F(ab′)2 reagents were used for Ig detection (each commercially absorbed against human serum proteins): donkey anti-goat IgG (code 705-066-147; Jackson Immunoresearch), donkey anti-rabbit IgG (code 711-066-152; Jackson Immunoresearch), horse anti-mouse IgG (BA-2000; Vector Laboratories), or goat anti-mouse IgM (μ-chain–specific; code 115-066-075; Jackson Immunoresearch).

    Article Title: Distinct Projection Targets Define Subpopulations of Mouse Brainstem Vagal Neurons that Express the Autism-Associated MET Receptor Tyrosine Kinase
    Article Snippet: Here, sections were stored at −80°C until used for MET and ChAT immunohistochemical processing, as previously described ( ). .. Sections were then washed several times in Blotto, and incubated for 1 hr at room temperature in Blotto containing Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Mouse IgG (Jackson ImmunoResearch Labs Cat# 715-066-150, RRID:AB_2340787) or Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs Cat# 705-066-147, RRID:AB_2340398).

    Article Title: Reduced infarct size in neuroglobin-null mice after experimental stroke in vivo
    Article Snippet: Antibody validation IHC was performed as described above using the following primary antibodies: Ngb was detected with either an in-house made polyclonal rabbit antibody (code# 4836/5, in 1:100.000 dilution), polyclonal rabbit antibody against amino acid 55–70 of human Ngb (code# N7162, Sigma-Aldrich, in 1:500 dilution) or polyclonal goat antibody against the an internal region of human Ngb (code# sc-22001, Santa Cruz Biotechnology, in 1:500 dilution). .. The primary antibodies were visualized by a (Fab)2 fragment of a donkey anti-rabbit antibody or goat conjugated to biotin (Jackson Immunoresearch Laboratories, Baltimore, PA, USA, code# 706-066-152 or 705-066-147, in 1:2000 dilution) in combination with ABC (Vector labs, Burlingame, CA, USA), followed by 0.05% DAB.

    Immunoprecipitation:

    Article Title: Endothelial CaMKII as a regulator of eNOS activity and NO-mediated vasoreactivity
    Article Snippet: Antibodies directed to eNOS (#9586) (used for immunoprecipitation), GAPDH (#2178), calmodulin (#4830), phosphoThr-287 CaMKII (#12716), phosphoSer-1179 in bovine eNOS (#9571), phosphoThr-497 in bovine eNOS (pThr-495 in human) (#9574) and anti-mouse IgG-HRP (#7076S) were purchased from Cell Signaling Technology. .. Goat anti-rabbit IgG (111-066-003) and donkey anti-goat IgG (705-066-147) biotinylated antibodies and normal rabbit and normal mouse IgG were from Jackson Immuno Research.

    Affinity Purification:

    Article Title: High Mobility Group 1 Protein (Hmg-1) Stimulates Proinflammatory Cytokine Synthesis in Human Monocytes
    Article Snippet: TNF-α (AF-210), IL-1α (AF-200), IL-1RA (AF-280), IL-6 (AF-206), IL-8 (AF-208), IL-12 (AF-219), macrophage inflammatory protein (MIP)-1α (BAF-270), and MIP-1β (BAF-271) were antigen affinity-purified goat IgG antibodies from R & D Systems; TNF-α (mAb1+mAb11, mouse IgG1 mAb) was from BD PharMingen, IL-1β (2.D.8+1437.96-15 mouse IgG1 mAb) was from Immunokontakt). .. Secondary antibodies for immunocytochemistry included Fab2 -fragmented biotinylated donkey anti–goat IgG (cat. no. 705-066-147; Jackson ImmunoResearch Laboratories) diluted 1:1,000; biotinylated goat anti–mouse IgG1 (cat. no. M32115; Caltag Laboratories) diluted 1:600; or biotinylated goat anti–rat IgG (cat. no. BA9400; Vector Laboratories) diluted 1:500.

    Article Title: TLR-9 and IL-15 Synergy Promotes the In Vitro Clonal Expansion of Chronic Lymphocytic Leukemia B Cells
    Article Snippet: .. The following secondary biotin-conjugated, affinity-purified F(ab′)2 reagents were used for Ig detection (each commercially absorbed against human serum proteins): donkey anti-goat IgG (code 705-066-147; Jackson Immunoresearch), donkey anti-rabbit IgG (code 711-066-152; Jackson Immunoresearch), horse anti-mouse IgG (BA-2000; Vector Laboratories), or goat anti-mouse IgM (μ-chain–specific; code 115-066-075; Jackson Immunoresearch). .. Each secondary Ab was followed by treatment with Vectastain ABC kit (Elite, pk6100-standard; Vector Laboratories) and detection achieved with the ImmPACT DAB Peroxidase substrate kit (Sk 4105; Vector laboratories).

    Binding Assay:

    Article Title: YKL-40 Is Differentially Expressed in Human Embryonic Stem Cells and in Cell Progeny of the Three Germ Layers
    Article Snippet: Nonspecific binding was inhibited by incubation for 30 min with 10% donkey serum (017-000-1210; Jackson ImmunoResearch). .. Sections were then incubated with HNF-3β for 1½ hr at room temperature and then in biotin-SP-conjugated F(ab′)2 fragment donkey anti-goat (705-066-147; Jackson ImmunoResearch) followed by streptavidin-conjugated Dylight 594 (SA5594; Vector Laboratories) before being coverslipped.

    Immunofluorescence:

    Article Title: The Immune-Modulating Cytokine and Endogenous Alarmin Interleukin-33 Is Upregulated in Skin Exposed to Inflammatory UVB Radiation
    Article Snippet: Paragraph title: Histology and Immunofluorescence ... After this, the TBS-washed slides were incubated for 1 hour at 37°C with a 1/200 biotin-conjugated donkey anti-goat IgG secondary antibody (705-066-147; Jackson Immunoresearch, West Grove, PA).

    Avidin-Biotin Assay:

    Article Title: Downstream Toll-like receptor signaling mediates adaptor-specific cytokine expression following focal cerebral ischemia
    Article Snippet: Immunoreactivity with primary antibody was visualized with avidin-biotin staining (ABC kit; Vector Laboratories, Burlingame, CA, USA) and diaminobenzene (DAB) staining (Vector Laboratories). .. Secondary antibodies included biotin-conjugated donkey anti-rat (cat. no. 712-066-153) and donkey anti-goat (cat. no. 705-066-147) at 1:3,000 concentrations (both from Jackson Immunoresearch Laboratories, West Grove, PA, USA) for anti-Ly6B.2 and anti-CD45, respectively.

    Article Title: TLR-9 and IL-15 Synergy Promotes the In Vitro Clonal Expansion of Chronic Lymphocytic Leukemia B Cells
    Article Snippet: The tissues were preblocked with 25% normal serum matching the secondary detection Ab’s species and further subjected to avidin/biotin blocking (kit from Vector Labs) before incubation at 4°C overnight in a humidified chamber with primary Ab: goat anti-human IL-15 (AF 315; R & D Systems), mouse anti-human Pax-5 (IS650, clone DAK; Dako), or mouse anti-human Ki-67 (N1633, clone Mib 1; Dako) at the recommended dilutions along with appropriate isotype controls. .. The following secondary biotin-conjugated, affinity-purified F(ab′)2 reagents were used for Ig detection (each commercially absorbed against human serum proteins): donkey anti-goat IgG (code 705-066-147; Jackson Immunoresearch), donkey anti-rabbit IgG (code 711-066-152; Jackson Immunoresearch), horse anti-mouse IgG (BA-2000; Vector Laboratories), or goat anti-mouse IgM (μ-chain–specific; code 115-066-075; Jackson Immunoresearch).

    Article Title: Increased Feeding and Food Hoarding following Food Deprivation Are Associated with Activation of Dopamine and Orexin Neurons in Male Brandt's Voles
    Article Snippet: Sections were rinsed and incubated in avidin-biotin complex (Vector) in 0.1 M PBS for 90 min, and rinsed in 0.1 M PBS. .. The same protocol outlined above was used except that the primary antibody for orexin labeling was goat anti-orexin (sc-8070; 1∶2000; Santa Cruz Biotechnology) and the secondary antibody was donkey anti-goat IgG (705-066-147; 1∶500; Jackson ImmunoResearch, West Grove, PA).

    Labeling:

    Article Title: The Immune-Modulating Cytokine and Endogenous Alarmin Interleukin-33 Is Upregulated in Skin Exposed to Inflammatory UVB Radiation
    Article Snippet: After this, the TBS-washed slides were incubated for 1 hour at 37°C with a 1/200 biotin-conjugated donkey anti-goat IgG secondary antibody (705-066-147; Jackson Immunoresearch, West Grove, PA). .. In some experiments, the fluorescently labeled sections were photographed immediately before the removal of the coverslip and subsequent staining of mast cells using toluidine blue as we have described.

    Article Title: Increased Feeding and Food Hoarding following Food Deprivation Are Associated with Activation of Dopamine and Orexin Neurons in Male Brandt's Voles
    Article Snippet: .. The same protocol outlined above was used except that the primary antibody for orexin labeling was goat anti-orexin (sc-8070; 1∶2000; Santa Cruz Biotechnology) and the secondary antibody was donkey anti-goat IgG (705-066-147; 1∶500; Jackson ImmunoResearch, West Grove, PA). .. The specificity of antibodies was assessed in rats and voles previously – , and also corroborated with control sections that were stained with preimmune rabbit (for c-Fos and TH) or goat (for orexin) serum; and control sections in which either the primary antibody, secondary antibody or the ABC complex was omitted.

    Purification:

    Article Title: The Immune-Modulating Cytokine and Endogenous Alarmin Interleukin-33 Is Upregulated in Skin Exposed to Inflammatory UVB Radiation
    Article Snippet: Slides were then incubated for 18 hours at 4°C with 10 μg/mL goat anti-mouse IL-33 (AF3626; R & D Systems, Minneapolis, MN), goat anti-human IL-33 (AF3625; R & D Systems), or purified goat IgG (AB-108-C; R & D Systems). .. After this, the TBS-washed slides were incubated for 1 hour at 37°C with a 1/200 biotin-conjugated donkey anti-goat IgG secondary antibody (705-066-147; Jackson Immunoresearch, West Grove, PA).

    Blocking Assay:

    Article Title: Downstream Toll-like receptor signaling mediates adaptor-specific cytokine expression following focal cerebral ischemia
    Article Snippet: Bovine serum albumin (Sigma-Aldrich) and 10% normal serum from the originating species of the secondary antibody were used for blocking. .. Secondary antibodies included biotin-conjugated donkey anti-rat (cat. no. 712-066-153) and donkey anti-goat (cat. no. 705-066-147) at 1:3,000 concentrations (both from Jackson Immunoresearch Laboratories, West Grove, PA, USA) for anti-Ly6B.2 and anti-CD45, respectively.

    Article Title: Distinct Projection Targets Define Subpopulations of Mouse Brainstem Vagal Neurons that Express the Autism-Associated MET Receptor Tyrosine Kinase
    Article Snippet: Briefly, slides were incubated for 2 hr in blocking buffer containing 5% normal donkey serum (Jackson ImmunoResearch, West Grove, PA) and 0.3% triton-X100 (Sigma, St. Louis, MO) in PBS, followed by incubation overnight in PBS containing 2% normal donkey serum, 0.3% triton-X100, and primary antibodies directed against one or more of the following antigens: Cocaine- and amphetamine-regulated transcript (CART) (Phoenix Pharmaceuticals Cat# H-003-62, RRID:AB_2313614), Choline Acetyltransferase (ChAT) (Millipore Cat# AB144P, RRID:AB_2079751), Green Fluorescent Protein (GFP) (Abcam Cat# ab13970, RRID:AB_300798), MET receptor tyrosine kinase (MET) (R and D Systems Cat# AF527, RRID:AB_355414), Neurofilament (DSHB Cat# 2H3, RRID:AB_531793), or Tyrosine Hydroxylase (TH) (Millipore Cat# AB152, RRID:AB_390204). .. After several rinses in PBS, tissue was then incubated for 2 hr at room temperature in PBS containing 2% normal donkey serum, 0.3% triton-X100, and one or more of the following species-specific secondary antibodies: Alexa Fluor® 488 AffiniPure F(ab′)2 Fragment Donkey Anti-Chicken IgG (Jackson ImmunoResearch Labs Cat# 703-546-155, RRID:AB_2340376), Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs Cat# 705-066-147, RRID:AB_2340398), DyLight 549 Streptavidin (Vector Laboratories Cat# SA-5549, RRID:AB_2336408), Alexa Fluor® 647 Donkey anti-Goat IgG (Thermo Fisher Scientific Cat# A-21447, RRID:AB_2535864), Cy3-AffiniPure Donkey Anti-Rabbit IgG, (Jackson ImmunoResearch Labs Cat# 711-165-152, RRID:AB_2307443), Alexa Fluor® 647 Donkey anti-Rabbit IgG, (Thermo Fisher Scientific Cat# A-31573, RRID:AB_2536183), and/or Alexa Fluor® 647 Donkey anti-Mouse IgG (Thermo Fisher Scientific Cat# A-31571, RRID:AB_2536181).

    Article Title: TLR-9 and IL-15 Synergy Promotes the In Vitro Clonal Expansion of Chronic Lymphocytic Leukemia B Cells
    Article Snippet: The tissues were preblocked with 25% normal serum matching the secondary detection Ab’s species and further subjected to avidin/biotin blocking (kit from Vector Labs) before incubation at 4°C overnight in a humidified chamber with primary Ab: goat anti-human IL-15 (AF 315; R & D Systems), mouse anti-human Pax-5 (IS650, clone DAK; Dako), or mouse anti-human Ki-67 (N1633, clone Mib 1; Dako) at the recommended dilutions along with appropriate isotype controls. .. The following secondary biotin-conjugated, affinity-purified F(ab′)2 reagents were used for Ig detection (each commercially absorbed against human serum proteins): donkey anti-goat IgG (code 705-066-147; Jackson Immunoresearch), donkey anti-rabbit IgG (code 711-066-152; Jackson Immunoresearch), horse anti-mouse IgG (BA-2000; Vector Laboratories), or goat anti-mouse IgM (μ-chain–specific; code 115-066-075; Jackson Immunoresearch).

    Article Title: Distinct Projection Targets Define Subpopulations of Mouse Brainstem Vagal Neurons that Express the Autism-Associated MET Receptor Tyrosine Kinase
    Article Snippet: Sections were incubated overnight at 4°C in blocking solution containing primary antibodies directed against MET (Cell Signaling Technology Cat# 3127, RRID:AB_331361) or ChAT (Millipore Cat# AB144P, RRID:AB_2079751). .. Sections were then washed several times in Blotto, and incubated for 1 hr at room temperature in Blotto containing Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Mouse IgG (Jackson ImmunoResearch Labs Cat# 715-066-150, RRID:AB_2340787) or Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs Cat# 705-066-147, RRID:AB_2340398).

    Article Title: The Immune-Modulating Cytokine and Endogenous Alarmin Interleukin-33 Is Upregulated in Skin Exposed to Inflammatory UVB Radiation
    Article Snippet: Slides were treated with a biotin blocking kit (X0590; Dako, Carpinteria, CA) as per the manufacturer's instructions before blocking other proteins with 5% normal equine serum for 1 hour at 25°C. .. After this, the TBS-washed slides were incubated for 1 hour at 37°C with a 1/200 biotin-conjugated donkey anti-goat IgG secondary antibody (705-066-147; Jackson Immunoresearch, West Grove, PA).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Specific Connectivity and Unique Molecular Identity of MET Receptor Tyrosine Kinase-Expressing Serotonergic Neurons in the Caudal Dorsal Raphe Nuclei
    Article Snippet: .. Secondary antibodies used were as follows: Alexa 647 AffiniPure F(ab′)2 Fragment Donkey Anti-Chicken IgG (Jackson ImmunoResearch Labs, Cat# 703-546-155), Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs, Cat# 705-066-147), DyLight 549 Streptavidin (Jackson ImmunoResearch Labs, Cat016-580-084), Alexa 647 Donkey anti-Rabbit IgG (Thermo Fisher Scientific Cat# A-31573). ..

    Mouse Assay:

    Article Title: Downstream Toll-like receptor signaling mediates adaptor-specific cytokine expression following focal cerebral ischemia
    Article Snippet: Secondary antibodies included biotin-conjugated donkey anti-rat (cat. no. 712-066-153) and donkey anti-goat (cat. no. 705-066-147) at 1:3,000 concentrations (both from Jackson Immunoresearch Laboratories, West Grove, PA, USA) for anti-Ly6B.2 and anti-CD45, respectively. .. Primary antibody immunoreactivity in sections from WT mice were compared to those from MyD88−/− and TRIF-mutant mice brain sections.

    Article Title: Distinct Projection Targets Define Subpopulations of Mouse Brainstem Vagal Neurons that Express the Autism-Associated MET Receptor Tyrosine Kinase
    Article Snippet: Mice used on or after P7 were deeply anesthetized by intraperitoneal injection of ketamine: xylazine (100 mg/kg: 10 mg/kg, Henry Schein, Melville, NY) and perfused with 0.9% saline, followed by fixative (4% paraformaldehyde (Sigma, St. Louis, MO) in 0.1M phosphate buffered saline (PBS), pH 7.4). .. After several rinses in PBS, tissue was then incubated for 2 hr at room temperature in PBS containing 2% normal donkey serum, 0.3% triton-X100, and one or more of the following species-specific secondary antibodies: Alexa Fluor® 488 AffiniPure F(ab′)2 Fragment Donkey Anti-Chicken IgG (Jackson ImmunoResearch Labs Cat# 703-546-155, RRID:AB_2340376), Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs Cat# 705-066-147, RRID:AB_2340398), DyLight 549 Streptavidin (Vector Laboratories Cat# SA-5549, RRID:AB_2336408), Alexa Fluor® 647 Donkey anti-Goat IgG (Thermo Fisher Scientific Cat# A-21447, RRID:AB_2535864), Cy3-AffiniPure Donkey Anti-Rabbit IgG, (Jackson ImmunoResearch Labs Cat# 711-165-152, RRID:AB_2307443), Alexa Fluor® 647 Donkey anti-Rabbit IgG, (Thermo Fisher Scientific Cat# A-31573, RRID:AB_2536183), and/or Alexa Fluor® 647 Donkey anti-Mouse IgG (Thermo Fisher Scientific Cat# A-31571, RRID:AB_2536181).

    Injection:

    Article Title: Distinct Projection Targets Define Subpopulations of Mouse Brainstem Vagal Neurons that Express the Autism-Associated MET Receptor Tyrosine Kinase
    Article Snippet: Mice used on or after P7 were deeply anesthetized by intraperitoneal injection of ketamine: xylazine (100 mg/kg: 10 mg/kg, Henry Schein, Melville, NY) and perfused with 0.9% saline, followed by fixative (4% paraformaldehyde (Sigma, St. Louis, MO) in 0.1M phosphate buffered saline (PBS), pH 7.4). .. After several rinses in PBS, tissue was then incubated for 2 hr at room temperature in PBS containing 2% normal donkey serum, 0.3% triton-X100, and one or more of the following species-specific secondary antibodies: Alexa Fluor® 488 AffiniPure F(ab′)2 Fragment Donkey Anti-Chicken IgG (Jackson ImmunoResearch Labs Cat# 703-546-155, RRID:AB_2340376), Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs Cat# 705-066-147, RRID:AB_2340398), DyLight 549 Streptavidin (Vector Laboratories Cat# SA-5549, RRID:AB_2336408), Alexa Fluor® 647 Donkey anti-Goat IgG (Thermo Fisher Scientific Cat# A-21447, RRID:AB_2535864), Cy3-AffiniPure Donkey Anti-Rabbit IgG, (Jackson ImmunoResearch Labs Cat# 711-165-152, RRID:AB_2307443), Alexa Fluor® 647 Donkey anti-Rabbit IgG, (Thermo Fisher Scientific Cat# A-31573, RRID:AB_2536183), and/or Alexa Fluor® 647 Donkey anti-Mouse IgG (Thermo Fisher Scientific Cat# A-31571, RRID:AB_2536181).

    Plasmid Preparation:

    Article Title: TLR-9 and IL-15 Synergy Promotes the In Vitro Clonal Expansion of Chronic Lymphocytic Leukemia B Cells
    Article Snippet: The following secondary biotin-conjugated, affinity-purified F(ab′)2 reagents were used for Ig detection (each commercially absorbed against human serum proteins): donkey anti-goat IgG (code 705-066-147; Jackson Immunoresearch), donkey anti-rabbit IgG (code 711-066-152; Jackson Immunoresearch), horse anti-mouse IgG (BA-2000; Vector Laboratories), or goat anti-mouse IgM (μ-chain–specific; code 115-066-075; Jackson Immunoresearch). .. The following secondary biotin-conjugated, affinity-purified F(ab′)2 reagents were used for Ig detection (each commercially absorbed against human serum proteins): donkey anti-goat IgG (code 705-066-147; Jackson Immunoresearch), donkey anti-rabbit IgG (code 711-066-152; Jackson Immunoresearch), horse anti-mouse IgG (BA-2000; Vector Laboratories), or goat anti-mouse IgM (μ-chain–specific; code 115-066-075; Jackson Immunoresearch).

    Article Title: Distinct Projection Targets Define Subpopulations of Mouse Brainstem Vagal Neurons that Express the Autism-Associated MET Receptor Tyrosine Kinase
    Article Snippet: Sections were then washed several times in Blotto, and incubated for 1 hr at room temperature in Blotto containing Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Mouse IgG (Jackson ImmunoResearch Labs Cat# 715-066-150, RRID:AB_2340787) or Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs Cat# 705-066-147, RRID:AB_2340398). .. After washing in PBS, sections were incubated for 1 hr in Vectastain ABC Elite solution (Vector Labs, Burlingame, CA, RRID:AB_2336819), washed in PBS, and incubated for 4 minutes in ImmPACT DAB Peroxidase (HRP) Substrate (Vector Labs, Burlingame, CA, RRID:AB_2336520).

    Article Title: Endothelial CaMKII as a regulator of eNOS activity and NO-mediated vasoreactivity
    Article Snippet: Goat anti-rabbit IgG (111-066-003) and donkey anti-goat IgG (705-066-147) biotinylated antibodies and normal rabbit and normal mouse IgG were from Jackson Immuno Research. .. Goat anti-rabbit IgG (111-066-003) and donkey anti-goat IgG (705-066-147) biotinylated antibodies and normal rabbit and normal mouse IgG were from Jackson Immuno Research.

    Article Title: The Immune-Modulating Cytokine and Endogenous Alarmin Interleukin-33 Is Upregulated in Skin Exposed to Inflammatory UVB Radiation
    Article Snippet: After this, the TBS-washed slides were incubated for 1 hour at 37°C with a 1/200 biotin-conjugated donkey anti-goat IgG secondary antibody (705-066-147; Jackson Immunoresearch, West Grove, PA). .. After this, the TBS-washed slides were incubated for 1 hour at 37°C with a 1/200 biotin-conjugated donkey anti-goat IgG secondary antibody (705-066-147; Jackson Immunoresearch, West Grove, PA).

    Article Title: Increased Feeding and Food Hoarding following Food Deprivation Are Associated with Activation of Dopamine and Orexin Neurons in Male Brandt's Voles
    Article Snippet: Cytoplasmic labeling was immunoreactively stained by using a Vector DAB Kit in 5 ml ddH2 O for 3 min, revealing brown cytoplasmic staining. .. The same protocol outlined above was used except that the primary antibody for orexin labeling was goat anti-orexin (sc-8070; 1∶2000; Santa Cruz Biotechnology) and the secondary antibody was donkey anti-goat IgG (705-066-147; 1∶500; Jackson ImmunoResearch, West Grove, PA).

    Microscopy:

    Article Title: Specific Connectivity and Unique Molecular Identity of MET Receptor Tyrosine Kinase-Expressing Serotonergic Neurons in the Caudal Dorsal Raphe Nuclei
    Article Snippet: 20μm sections were cut using a cryostat, mounted onto Superfrost Plus microscope slides (VWR, Radnor, PA), and processed for immunostaining as described , . .. Secondary antibodies used were as follows: Alexa 647 AffiniPure F(ab′)2 Fragment Donkey Anti-Chicken IgG (Jackson ImmunoResearch Labs, Cat# 703-546-155), Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs, Cat# 705-066-147), DyLight 549 Streptavidin (Jackson ImmunoResearch Labs, Cat#016-580-084), Alexa 647 Donkey anti-Rabbit IgG (Thermo Fisher Scientific Cat# A-31573).

    Article Title: Distinct Projection Targets Define Subpopulations of Mouse Brainstem Vagal Neurons that Express the Autism-Associated MET Receptor Tyrosine Kinase
    Article Snippet: Four series of 20μm sections were cut using a cryostat, mounted onto superfrost plus microscope slides (VWR, Radnor, PA), and processed for immunostaining. .. After several rinses in PBS, tissue was then incubated for 2 hr at room temperature in PBS containing 2% normal donkey serum, 0.3% triton-X100, and one or more of the following species-specific secondary antibodies: Alexa Fluor® 488 AffiniPure F(ab′)2 Fragment Donkey Anti-Chicken IgG (Jackson ImmunoResearch Labs Cat# 703-546-155, RRID:AB_2340376), Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs Cat# 705-066-147, RRID:AB_2340398), DyLight 549 Streptavidin (Vector Laboratories Cat# SA-5549, RRID:AB_2336408), Alexa Fluor® 647 Donkey anti-Goat IgG (Thermo Fisher Scientific Cat# A-21447, RRID:AB_2535864), Cy3-AffiniPure Donkey Anti-Rabbit IgG, (Jackson ImmunoResearch Labs Cat# 711-165-152, RRID:AB_2307443), Alexa Fluor® 647 Donkey anti-Rabbit IgG, (Thermo Fisher Scientific Cat# A-31573, RRID:AB_2536183), and/or Alexa Fluor® 647 Donkey anti-Mouse IgG (Thermo Fisher Scientific Cat# A-31571, RRID:AB_2536181).

    Article Title: TLR-9 and IL-15 Synergy Promotes the In Vitro Clonal Expansion of Chronic Lymphocytic Leukemia B Cells
    Article Snippet: The following secondary biotin-conjugated, affinity-purified F(ab′)2 reagents were used for Ig detection (each commercially absorbed against human serum proteins): donkey anti-goat IgG (code 705-066-147; Jackson Immunoresearch), donkey anti-rabbit IgG (code 711-066-152; Jackson Immunoresearch), horse anti-mouse IgG (BA-2000; Vector Laboratories), or goat anti-mouse IgM (μ-chain–specific; code 115-066-075; Jackson Immunoresearch). .. Most images were photographed at final magnifications of 100×, 200×, or 600× (the latter with a 60×/1.25 Oil Iris ∞/0.17) using an Olympus BX40 phase-contrast microscope and Olympus DP20 camera.

    Concentration Assay:

    Article Title: High Mobility Group 1 Protein (Hmg-1) Stimulates Proinflammatory Cytokine Synthesis in Human Monocytes
    Article Snippet: The cytokine-specific antibodies were used at a final concentration of 2–5 μg/ml. .. Secondary antibodies for immunocytochemistry included Fab2 -fragmented biotinylated donkey anti–goat IgG (cat. no. 705-066-147; Jackson ImmunoResearch Laboratories) diluted 1:1,000; biotinylated goat anti–mouse IgG1 (cat. no. M32115; Caltag Laboratories) diluted 1:600; or biotinylated goat anti–rat IgG (cat. no. BA9400; Vector Laboratories) diluted 1:500.

    Marker:

    Article Title: Distinct Projection Targets Define Subpopulations of Mouse Brainstem Vagal Neurons that Express the Autism-Associated MET Receptor Tyrosine Kinase
    Article Snippet: Sections were then washed several times in Blotto, and incubated for 1 hr at room temperature in Blotto containing Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Mouse IgG (Jackson ImmunoResearch Labs Cat# 715-066-150, RRID:AB_2340787) or Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs Cat# 705-066-147, RRID:AB_2340398). .. As a reference marker for neuronal subpopulations in the rhesus macaque brainstem, ChAT immunostaining was performed to localize cholinergic neurons in sections proximal to the GD151 brainstem sections used for MET immunostaining.

    Staining:

    Article Title: Downstream Toll-like receptor signaling mediates adaptor-specific cytokine expression following focal cerebral ischemia
    Article Snippet: Immunoreactivity with primary antibody was visualized with avidin-biotin staining (ABC kit; Vector Laboratories, Burlingame, CA, USA) and diaminobenzene (DAB) staining (Vector Laboratories). .. Secondary antibodies included biotin-conjugated donkey anti-rat (cat. no. 712-066-153) and donkey anti-goat (cat. no. 705-066-147) at 1:3,000 concentrations (both from Jackson Immunoresearch Laboratories, West Grove, PA, USA) for anti-Ly6B.2 and anti-CD45, respectively.

    Article Title: Distinct Projection Targets Define Subpopulations of Mouse Brainstem Vagal Neurons that Express the Autism-Associated MET Receptor Tyrosine Kinase
    Article Snippet: Paragraph title: 3,3′-Diaminobenzidine (DAB) Staining ... Sections were then washed several times in Blotto, and incubated for 1 hr at room temperature in Blotto containing Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Mouse IgG (Jackson ImmunoResearch Labs Cat# 715-066-150, RRID:AB_2340787) or Biotin-SP-AffiniPure F(ab′)2 Fragment Donkey Anti-Goat IgG (Jackson ImmunoResearch Labs Cat# 705-066-147, RRID:AB_2340398).

    Article Title: The Immune-Modulating Cytokine and Endogenous Alarmin Interleukin-33 Is Upregulated in Skin Exposed to Inflammatory UVB Radiation
    Article Snippet: After this, the TBS-washed slides were incubated for 1 hour at 37°C with a 1/200 biotin-conjugated donkey anti-goat IgG secondary antibody (705-066-147; Jackson Immunoresearch, West Grove, PA). .. In some experiments, the fluorescently labeled sections were photographed immediately before the removal of the coverslip and subsequent staining of mast cells using toluidine blue as we have described.

    Article Title: Increased Feeding and Food Hoarding following Food Deprivation Are Associated with Activation of Dopamine and Orexin Neurons in Male Brandt's Voles
    Article Snippet: Cytoplasmic labeling was immunoreactively stained by using a Vector DAB Kit in 5 ml ddH2 O for 3 min, revealing brown cytoplasmic staining. .. The same protocol outlined above was used except that the primary antibody for orexin labeling was goat anti-orexin (sc-8070; 1∶2000; Santa Cruz Biotechnology) and the secondary antibody was donkey anti-goat IgG (705-066-147; 1∶500; Jackson ImmunoResearch, West Grove, PA).

    Article Title: YKL-40 Is Differentially Expressed in Human Embryonic Stem Cells and in Cell Progeny of the Three Germ Layers
    Article Snippet: After staining for YKL-40, the sections were subjected to antigen retrieval in TEG buffer. .. Sections were then incubated with HNF-3β for 1½ hr at room temperature and then in biotin-SP-conjugated F(ab′)2 fragment donkey anti-goat (705-066-147; Jackson ImmunoResearch) followed by streptavidin-conjugated Dylight 594 (SA5594; Vector Laboratories) before being coverslipped.

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