PA5-101099

antibody from Invitrogen Antibodies

Targeting: ITPKB IP3-3KB, IP3KB

Provider product page for PA5-101099

Western blot

Immunocytochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [1]
Comments [0]
Validations
Immunocytochemistry [1]
Other assay [4]

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Product number: PA5-101099 - Provider product page
Provider: Invitrogen Antibodies
Product name: ITPKB Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: Antibody detects endogenous levels of total IP3KB.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 1 mg/mL
Storage: -20°C

ITPKB protein structure - PA5-101099 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 3 miR-711 targets and negatively regulates Itpkb. (A) The volcano expression graph of the expression profile GSE5281. Red dots indicate upregulated genes and green dots indicate downregulated genes. (B) TargetScan prediction of downstream genes of miR-711 and Venn diagram of the top 15 genes of expression profile GSE5281. The four genes in the intersection are ERBB2IP, SEMA4C, Itpkb, and DTNA. (C) PPI network diagram of the four intersections and their related genes constructed by String. A higher gene core degree corresponds to a darker circle color, whereas a lower core degree corresponds to a bluer color. (D) Itpkb expression box plot drawn based on the data of expression profile GSE5281. The blue box on the left indicates the expression in normal samples, and the red box on the right indicates the expression in AD samples. (E) The possible binding site of miR-711 to Itpkb predicted by TargetScan. (F) Dual-luciferase reporter gene assay to verify whether miR-711 can target Itpkb. (G) RT-qPCR to detect the expression of miR-711 in BV2 cells in each group. (H) Western blot analysis to detect the expression of Itpkb normalized to GAPDH in BV2 cells in each group. * p < 0.05 versus the NC mimic group; # p < 0.05 versus the NC inhibitor group. The measurement data were summarized as mean +- standard deviation. Unpaired t -test was used for two-group comparisons. One-way ANOVA was carried out for comparisons among multiple groups followed by Tukey's post hoc test. Cellula

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 4 miR-711 mediates anti-inflammatory pathway of microglia and M2-like phenotype by targeting Itpkb. (A) RT-qPCR to detect the expression of miR-711 and Itpkb in each group of BV2 cells. (B) Western blot assay to detect the expression of Itpkb normalized to GAPDH in each group of BV2 cells. (C) ELISA to detect the expression of inflammatory factor TNF-alpha and anti-inflammatory factor IL-10 in BV2 cell culture medium. (D) Flow cytometry to detect the content of M2 phenotype and M1 phenotype microglia in BV2 cells, with the ratio calculated. * p < 0.05 versus the NC mimic-OE-NC group; # p < 0.05 versus the NC mimic-OE-Itpkb group. The measurement data were summarized as mean +- standard deviation. One-way ANOVA was carried out for comparisons among multiple groups followed by Tukey's post hoc test. Cellular experiment was repeated three times independently.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 5 miR-711 mediates Tau protein hyperphosphorylation via Itpkb pathway. (A) RT-qPCR detection of Itpkb expression in each group of BV2 cells. (B) Western blot detection of Itpkb, p-Tau and Tau expression normalized to GAPDH in each group of BV2 cells. (C) RT-qPCR detection of miR-711 and Itpkb expression in each group of BV2 cells. (D) Western blot detection of the expression of Itpkb, p-Tau and Tau normalized to GAPDH in BV2 cells of each group. * p < 0.05 versus the NC mimic-OE-NC group or DMSO-OE-NC group; # p < 0.05 versus the DMSO-OE-Itpkb group or NC mimic-OE-Itpkb group. The measurement data were summarized as mean +- standard deviation. One-way ANOVA was carried out for comparisons among multiple groups followed by Tukey's post hoc test. Cellular experiment was repeated three times independently.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 6 miR-711-loaded microglia-derived EVs alleviate inflammation, neurodegenerative changes and cognitive dysfunction in mice by targeting Itpkb. (A) RT-qPCR to detect the expression of miR-711 and Itpkb in the brain tissues of mice in each group. (B) Western blot assay to detect the expression of p-Tau and Itpkb protein in mouse brain tissues, normalized to GAPDH. (C) ELISA to detect the expression of inflammatory factor TNF-alpha and anti-inflammatory factor IL-10 in mouse serum. (D ) Flow cytometry to detect the content of M2 phenotype and M1 phenotype microglia in mouse brain tissue, with the ratio calculated. (E) After the model was successfully constructed, the mice were scored for neurological deficits on days 1, 3, 7, 14, 21, and 28. (F) Quantitative analysis of the Morris water maze escape latency of the mice in each group during 4 days of training. (G) Quantitative analysis of the number of times that the mice of each group crossed the target platform. (H) Quantitative analysis of the percentage of time spent by the mice in the target quadrant. (I) Quantitative analysis of the average swimming speed of mice in each group. * p < 0.05 versus the NC mimic-EVs + OE-NC group; # p < 0.05 versus the NC mimic-EVs + OE-Itpkb group. The measurement data were summarized as mean +- standard deviation. One-way ANOVA was carried out for comparisons among multiple groups followed by Tukey's post hoc test. Repeated measures ANOVA with Bonferroni post hoc test was employed to co