PA1-26938

antibody from Invitrogen Antibodies

Targeting: INS IDDM1, IDDM2

Provider product page for PA1-26938

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [20]
Comments [0]
Validations
Immunohistochemistry [1]
Other assay [9]

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Product number: PA1-26938 - Provider product page
Provider: Invitrogen Antibodies
Product name: Insulin Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Other
Description: This antibody stains beta(b) cells of the Langerhan's islets in human pancreas. IHC Note: prolonged fixation in buffered formalin can destroy the epitope.
Reactivity: Human, Mouse
Host: Guinea Pig
Isotype: IgG
Vial size: 500 µL
Concentration: 0.2 mg/mL
Storage: 4° C, do not freeze

INS protein structure - PA1-26938 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 Enhancer activity assessment and Scrt1 function. ( a ) Trn5 integrations on Mafb, Syt4, Neurod1 and two Pax6 accessible sites. ( b ) Scheme of pGL3 vector used for the luciferase assay. ( c ) Luciferase activity was measured in INS 832/13 cells transfected with an empty pGL3 vector (Ctrl) or a pGL3 vector containing an enhancer region for the indicated gene ( Mafb, NeuroD1 , Pax6 and Syt4 ). Results are expressed as fold change versus control. ( d ) Gene expression in P10 and adult rat islets were measured by qPCR and normalized to the housekeeping gene Hprt1 . Syt4 gene expression is available in Fig. 3 . * p < 0.05, ** p < 0.01 by Student''s t-test or by one-way Anova, Dunnett''s post-hoc test. ( e - f ) Scrt1 expression was measured by qPCR and normalized to Hprt1 housekeeping gene levels. ( f - j ) Dispersed adult rat islet cells were transfected with a control siRNA (siCtl) or siRNAs directed against Scrt1 (siScrt1). Experiments were performed 48 h post-transfection. Insulin release in response to 2 or 20 mM glucose ( g,h ) insulin content were determined by ELISA n = 3. ( i ) Apoptosis of insulin-positive cells was assessed using Tunel assay in basal (NT) condition or in response to a mix (cyt mix) of pro-inflammatory cytokines (IL-1beta, TNF-alpha and IFN-gamma) n = 5. ( j ) The fraction of proliferative insulin-positive cells was determined by BrdU incorporation in basal (NT) or stimulated (prolactin, PRL) conditions, n = 6. * p < 0.05, ** p < 0.01 by Student

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 VSG distinctly improves the identity and function of beta-cells . A,B , UMAP plots of 35,941 endocrine cells from Sham, VSG and PF. Colours in (a) indicate the five main endocrine cell clusters and in (b) distinguish beta-cells specifically according to the three groups. C , Abstracted graph of cluster connectivity assessing transcriptomic similarity between VSG-, PF-, Sham- and WT-derived beta-cells, inferred using PAGA. PAGA is based on a statistical method that measures relatedness between single-cell clusters, with edge weights indicating link significance. D,E , Gene expression for key genes involved in beta-cell identity and function changed upon VSG. (d) Dot plot showing that VSG-derived beta-cells upregulated essential maturity markers and downregulated dedifferentiation markers. (e) Dot plot showing genes critical for beta-cell function and metabolism, insulin processing and exocytosis. Color intensity shows normalized mean expression in a group; dot size indicates the percentage of cells that express the corresponding gene. Expression is scaled per gene. F-J , Immunohistochemical analysis of MafA, Slc2a2, Insulin, Aldh1a3, Slc5a10 and Pcsk1 in beta-cells of WT, Sham, PF and VSG mice at study end. Scale is 50 mum K , Volcano plots indicating differences in the number of genes that were differentially expressed when comparing beta-cells from Sham and VSG versus beta-cells from Sham and PF, obtained from differential analysis testing using limma-trend. Red dot

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 VSG rapidly expands beta-cell mass . A , Pancreas morphometry analysis reveals that VSG doubled the relative beta-cell area (WT, n = 6; Sham, n = 5, PF, n = 3; VSG, n = 3; WT versus VSG (p < 0.0001); Sham versus VSG (p < 0.0001), PF vs VSG (p = 0.0003)). B , Quantification of beta-cell proliferation at study end (WT, n = 6; Sham, n = 5, PF, n = 3; VSG, n = 3; WT versus VSG (p = 0.019), VSG versus Sham (p > 0.999), VSG versus PF (p = 0.156)). All data are presented as mean +- s.e.m and were analyzed by one-way ANOVA with Bonferroni's post-hoc test. C , Representative images of immuno-stainings against insulin, glucagon and EdU. Scale is 50 mum D , Dot plot showing expression of cell cycle-related genes in cycling and non-cycling beta-cells per group. Color intensity shows normalized mean expression in a cell cluster. Expression is scaled per gene. E , Doughnut plots indicating fraction of proliferating beta-cells per group (VSG-54/9252, PF-17/7389, Sham-14/4491). F , Dot plot showing expression of beta-cell identity genes in mature, dedifferentiated and proliferating beta-cells of VSG, implying the dedifferentiated signature of proliferating beta-cells. Expression is scaled per gene. G , Venn diagram showing uniquely upregulated beta-cell genes in VSG versus control groups. H , Top 10 unique VSG genes (estimated log (FC) > 0.25, B > 150). Log (FC) plotted for VSG versus Sham. I-J , Matrix plots showing expression of (i) immune-related and (j) pregnancy-related genes i

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 Impaired differentiation of pancreatic endocrine cells in Insm1 , Neurod1 , and Pax6 KO embryos. (A-D) Immunofluorescence labeling of pancreata from E18.5 Insm1 GFP -expressing embryos using antibodies against GFP (green) that marks pre-endocrine cells, and pancreatic hormones insulin (Ins, blue), and glucagon (Gcg, red). Compared with Insm1 +/- mice (A), mice lacking Insm1 (B), Neurod1 (C), and Pax6 (D) exhibit a decrease in total number of endocrine cells, altered endocrine cell morphology and numbers of hormone expressing cells. (E-H) Immunofluorescence labeling of pancreata from E18.5 embryos using antibodies against GFP (green), pancreatic polypeptide (Ppy, red), and somatostatin (Sst, blue) shows altered numbers of hormone expressing cells in Insm1 (F), Neurod1 (G), and Pax6 (H) KO mice. (I-L) Quantification of a percentage of hormone-positive cells among GFP-positive endocrine cells demonstrates defects in differentiation of cells positive for hormones: insulin (Ins) (I), glucagon (Gcg) (J), somatostatin (Sst) (K), and pancreatic polypeptide (Ppy) (L) in Insm1 , Neurod1 , and Pax6 KO embryonic pancreata in comparison with Insm1 +/- . Error bars indicate SEM ( n = 3); P -values were determined by one-way ANOVA test. Asterisks indicate P -values of *

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Increased number of CADM1 + CD45 + cells within the pancreatic islet during T1D. ( A ) Immunostaining of paraffin-embedded pancreata from individuals in the Non group for CADM1 (cyan), insulin (green), and GCG (red). Scale bar: 20 mum. ( B ) Immunostaining of paraffin-embedded pancreata from individuals in the Non, aAb + , and T1D groups for CADM1 (red) and CD45 (green). Scale bar: 50 mum. ( C ) Quantification of the number of CADM1 + CD45 + cells within the islet periphery ( n = 5 per group). ( D ) Quantification of the number of CD45 + adjacent to CADM1 + cells per area pancreas ( n = 5 per group). One-way ANOVA was performed using GraphPad Prism, version 7, software for comparisons of 3 groups. Post hoc statistical analyses were performed using the Tukey multiple comparisons test. Results are presented as mean +- SEM. * P < 0.05; *** P < 0.001. ad., adjacent.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Increased number of CD68 + cells adjacent to CADM1 + insulin + cells during T1D. ( A ) Immunostaining of paraffin-embedded pancreata from individuals in the Non, aAb + , and T1D groups for CADM1 (red), CD68 (green), and insulin (magenta). Scale bar: 50 mum. ( B ) Quantification of the number of CD68 + cells within the islet boundary ( n = 5 per group). ( C ) Quantification of the number of CD68 + cells at the islet periphery per islet area ( n = 5 per group). ( D ) Quantification of the number of CD68 + cells within the islet boundary ( n = 5 per group). ( E ) Quantification of the number of CADM1 + CD68 + cells within the islet boundary ( n = 5 per group). One-way ANOVA was performed using GraphPad Prism, version 7, software for comparisons of 3 groups. Post hoc statistical analyses were performed using Tukey's multiple comparisons test. Results are presented as mean +- SEM. * P < 0.05; ** P < 0.01; *** P < 0.001.