polyclonal goat igg af1180  (R&D Systems)

Journal: Diagnostic Pathology

Article Title: Establishment of monoclonal anti-human CD26 antibodies suitable for immunostaining of formalin-fixed tissue

doi: 10.1186/1746-1596-9-30

Figure Lengend Snippet: Flow cytometry analysis with novel anti-CD26 mAbs. A. Jurkat-CD26WT cells (red lines) or Jurkat parent cells (blue lines) were incubated with the hybridoma supernatant, and subsequently stained with PE-labeled anti-mouse Ig pAb, and analyzed by flow cytometry. B. Jurkat-CD26WT cells were incubated with the hybridoma supernatant (clone 1, 5, 11, 16, 18 or 19) or purified mouse anti-CD26 mAb (5F8) or commercial mouse anti-CD26 mAb (MBL, clone 44–4), and subsequently stained with PE-labeled anti-mouse Ig pAb, and analyzed by flow cytometry. The gray areas in each histogram show the data involving the isotype control. The mean fluorescence intensity (MFI) of each staining is shown. Data shown are repeated twice (A) and five times (B) with similar results.

Article Snippet: The sections were treated with 100 μl of hybridoma supernatants or purified mouse anti-human CD26 mAb or goat anti-human CD26 pAb for 2 hours at RT, and subsequently treated with HRP-conjugated horse anti-mouse Ig pAb or HRP-conjugated horse anti-goat IgG pAb (Vector Laboratories) for 30 min at RT.

Techniques: Flow Cytometry, Incubation, Staining, Labeling, Purification, Fluorescence

Journal: Diagnostic Pathology

Article Title: Establishment of monoclonal anti-human CD26 antibodies suitable for immunostaining of formalin-fixed tissue

doi: 10.1186/1746-1596-9-30

Figure Lengend Snippet: ELISA analysis with novel anti-CD26 mAbs. Non treated native soluble CD26 (sCD26) or urea treated denatured sCD26 was incubated with the hybridoma supernatant (clone 1, 5, 11, 16, 18 or 19) or purified mouse anti-CD26 mAb (5F8) or commercial mouse anti-CD26 mAb (MBL, clone 44–4) or purified goat anti-CD26 pAb (R&D Systems). The absorbance at 450 nm/570 nm was measured, and data are shown as mean ± S.E. from three independent experiments.

Article Snippet: The sections were treated with 100 μl of hybridoma supernatants or purified mouse anti-human CD26 mAb or goat anti-human CD26 pAb for 2 hours at RT, and subsequently treated with HRP-conjugated horse anti-mouse Ig pAb or HRP-conjugated horse anti-goat IgG pAb (Vector Laboratories) for 30 min at RT.

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Purification

Journal: Diagnostic Pathology

Article Title: Establishment of monoclonal anti-human CD26 antibodies suitable for immunostaining of formalin-fixed tissue

doi: 10.1186/1746-1596-9-30

Figure Lengend Snippet: Representative results of immunostaining with novel anti-CD26 mAbs. A. The tissue specimens of liver, kidney, prostate or two cases of malignant mesothelioma were stained with 100 μl of commercial mouse anti-human CD26 mAb supernatant (MBL, clone 44–4) (i), or 10 μg/ml of purified goat anti-human CD26 pAb (R&D Systems) (ii), or newly developed hybridoma supernatant (clone 18 (iii), clone 19 (iv) or clone 3 (v)). B . Malignant mesothelioma tissue specimens were stained with commercial goat anti-human CD26 pAb (R&D Systems) (i), or purified novel mouse anti-human CD26 mAbs (clone 18 (ii) or clone 19 (iii)) at the indicated concentrations of Abs in the presence or absence of sCD26. C. The tissue specimens of hepatocellular carcinoma, renal cell carcinoma, prostate adenocarcinoma, colon adenocarcinoma or lung adenocarcinoma were stained with 100 μg/ml of commercial goat anti-human CD26 pAb (R&D Systems) (i), or purified mouse anti-human CD26 mAbs (clone 18 (ii) or clone 19 (iii)). All specimens were counterstained with hematoxylin (original magnification, 200X).

Article Snippet: The sections were treated with 100 μl of hybridoma supernatants or purified mouse anti-human CD26 mAb or goat anti-human CD26 pAb for 2 hours at RT, and subsequently treated with HRP-conjugated horse anti-mouse Ig pAb or HRP-conjugated horse anti-goat IgG pAb (Vector Laboratories) for 30 min at RT.

Techniques: Immunostaining, Staining, Purification

Journal: Diagnostic Pathology

Article Title: Establishment of monoclonal anti-human CD26 antibodies suitable for immunostaining of formalin-fixed tissue

doi: 10.1186/1746-1596-9-30

Figure Lengend Snippet: Analysis of crossreactivity of novel anti-CD26 mAbs with humanized anti-CD26 mAb. Jurkat-CD26WT cells were pretreated with unlabeled humanized anti-CD26 mAb (YS110) or human control IgG, and then treated with the hybridoma supernatant (clone 1, 5, 11, 16, 18 or 19) or purified mouse anti-CD26 mAb (1F7 or 5F8), and subsequently stained with PE-labeled anti-mouse Ig pAb. For staining with humanized anti-CD26 mAb, cells were stained with Alexa Fluor 647-labeled YS110 after pretreatment with unlabeled YS110. Data were analyzed by flow cytometry, and the percentage of mean fluorescence intensity (MFI) after YS110 blocking to MFI after control IgG blocking is shown. Data shown are repeated twice with similar results.

Article Snippet: The sections were treated with 100 μl of hybridoma supernatants or purified mouse anti-human CD26 mAb or goat anti-human CD26 pAb for 2 hours at RT, and subsequently treated with HRP-conjugated horse anti-mouse Ig pAb or HRP-conjugated horse anti-goat IgG pAb (Vector Laboratories) for 30 min at RT.

Techniques: Purification, Staining, Labeling, Flow Cytometry, Fluorescence, Blocking Assay

Journal: Diagnostic Pathology

Article Title: Establishment of monoclonal anti-human CD26 antibodies suitable for immunostaining of formalin-fixed tissue

doi: 10.1186/1746-1596-9-30

Figure Lengend Snippet: Blocking experiment of novel anti-CD26 mAb binding to CD26. Jurkat-CD26WT cells were pretreated with unlabeled mouse anti-CD26 mAbs (4G8, 1F7, 5F8, 16D4B, or 9C11) (blue lines) or mouse IgG 1 isotype control (Contl. IgG) (red lines), and subsequently stained with PE-labeled anti-mouse Ig pAb (i) or Alexa Fluor 647-labeled anti-CD26 mAbs (clone 18 (ii) or clone 19 (iii) ), and analyzed by flow cytometry. The representative histograms of CD26 expression are shown, and the gray areas in each histogram show the data involving the isotype control. Data shown are repeated twice with similar results.

Article Snippet: The sections were treated with 100 μl of hybridoma supernatants or purified mouse anti-human CD26 mAb or goat anti-human CD26 pAb for 2 hours at RT, and subsequently treated with HRP-conjugated horse anti-mouse Ig pAb or HRP-conjugated horse anti-goat IgG pAb (Vector Laboratories) for 30 min at RT.

Techniques: Blocking Assay, Binding Assay, Staining, Labeling, Flow Cytometry, Expressing

Journal: Diagnostic Pathology

Article Title: Establishment of monoclonal anti-human CD26 antibodies suitable for immunostaining of formalin-fixed tissue

doi: 10.1186/1746-1596-9-30

Figure Lengend Snippet: Staining for CD26 expression on COS-7 cells transfected with CD26 deletion mutants by novel anti-CD26 mAbs. cDNA of deleted CD26 was cotransfected with GFP-expressing plasmid to COS-7 cells. After 24 h, the transfected cells were stained with Alexa Fluor 647-labeled anti-CD26 mAbs (YS110, clone 18 or clone 19) or isotype control, and analyzed by flow cytometry. Following gating for GFP positive cells among all acquired cells, the percentage of CD26 positive cells was analyzed. Data shown are repeated twice with similar results.

Article Snippet: The sections were treated with 100 μl of hybridoma supernatants or purified mouse anti-human CD26 mAb or goat anti-human CD26 pAb for 2 hours at RT, and subsequently treated with HRP-conjugated horse anti-mouse Ig pAb or HRP-conjugated horse anti-goat IgG pAb (Vector Laboratories) for 30 min at RT.

Techniques: Staining, Expressing, Transfection, Plasmid Preparation, Labeling, Flow Cytometry

Journal: JID Innovations

Article Title: Human keratinocytes exhibit limited potential for SARS-CoV-2 infection despite ACE2 and mature cathepsin L expression

doi: 10.1016/j.xjidi.2025.100447

Figure Lengend Snippet: Differential ACE2, TMPRSS2, NRP1, CTSL, CD147, AXL, and DPP4 protein expression. Whole-cell lysates of the indicated cells were analyzed by western blot using the following antibodies: ACE2 (mAb clone AC384), TMPRSS2 (mAb clone S20014A), NRP1 (mAb clone 14H4), CTSL (mAb clone 33/1), CD147 (mAb clone HIM6), AXL (polyclonal goat IgG, AF154), and DPP4 (polyclonal goat IgG, AF1180). ( a ) Representative immunoblots from 2 to 4 independent experiments. ( b ) Densitometric quantification normalized to actin, expressed as mean ± SEM from 2 to 4 independent experiments. CTSL, cathepsin L.

Article Snippet: Primary antibodies included anti-ACE2, clone AC384 (AdipoGen Life Science/Coger, Paris, France); anti-TMPRSS2, clone S20014A; anti-NRP1, clone 14H4; anti-CD147, clone HIM6; anti–keratin 10, rabbit polyclonal Poly19054 (BioLegend); anti-CTSL, clone 33/1 (eBioscience, Thermo Fisher Scientific); anti-AXL, polyclonal goat IgG AF154; and anti-DPP4, polyclonal goat IgG AF1180 (R&D Systems, Bio-Techne SAS, Noyal Châtillon, France).

Techniques: Expressing, Western Blot