- PA5-103060 - Invitrogen Antibodies BACH1 antibody

Du Y, Shi X, Li J, Jia Y
Experimental and therapeutic medicine 2021 Dec;22(6):1436

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of BACH1 in K562 cell lysate (left lane: treated with blocking peptide). Samples were incubated with BACH1 polyclonal antibody (Product # PA5-103060).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of BACH1 in RAW264.7 cell lysate. Samples were incubated with BACH1 polyclonal antibody (Product # PA5-103060).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of BACH1 in HepG2. Samples were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100, blocked with 10% serum (45 min at 25°C) incubated with BACH1 polyclonal antibody (Product # PA5-103060) using a dilution of 1:200 (1 hr, 37°C), and followed by goat anti-rabbit IgG Alexa Fluor 594 at a dilution of 1:600.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of paraffin-embedded BACH1 in human spleen tissue. Antigen retrieval was performed using citrate buffer. Samples were blocked with blocking buffer (1.5 hr, 22°C), incubated with BACH1 polyclonal antibody (Product # PA5-103060) using a dilution of 1:100 (1.5 hr, 22°C), followed by HRP conjugated goat anti-rabbit.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 miR-98-5p directly binds to the 3'-UTR of BACH1 to downregulate its expression. (A) Putative binding site of miR-98-5p in the 3'-UTR of BACH1. (B) BACH1 gene expression and (C) protein level in lupus nephritis tissue compared with control renal tissues. Human mesangial cells were treated with a miR-98-5p inhibitor, mimic and respective controls, after which (D) BACH1 gene and (E) protein expression were measured. (F) Human mesangial cells were co-transfected with either a miR-98-5p mimic and pmirGLO-BACH1-MUT or co-transfected with a miR-98-5p mimic and pmirGLO-BACH1-WT, after which relative luciferase activity was measured. All experiments were repeated three times and the data are expressed as the mean +- standard deviation. Data in panel B and F were analyzed by a paired Student's t-test. Data in panel D and E were analyzed by one-way ANOVA followed by a Tukey's post hoc test. ** P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 BACH1 promotes human mesangial cell proliferation. The effect of transfection with BACH1 siRNA on the levels of BACH1 expression at the (A) gene and (B) protein level were assessed. (C) Effect of transfection with BACH1 siRNA, BACH1-overexpression plasmid and BACH1-overexpression plasmid + BAY11-7082 (an inhibitor of NF-kappaB) on p-p65 and t-p65 protein levels. (D) Human mesangial cell proliferation was measured following transfection with the BACH1-overexpression plasmid, BACH1-overexpression plasmid + BAY11-7082, BACH1 siRNA and their respective controls. (E) Human mesangial cell secretion of TNF-alpha and IL-6 were measured after transfection with the BACH1-overexpression plasmid, BACH1-overexpression plasmid + BAY11-7082, BACH1 siRNA and their respective controls. All experiments were repeated three times and the data are expressed as the mean +- standard deviation. Data were analyzed by one-way ANOVA followed by a Tukey's post hoc test. ** P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Transfection with the miR-98-5p mimic inhibits the BACH1-overexpression induced proliferation of human mesangial cells. Effect of transfection with the BACH1-overexpression plasmid and co-transfection with the miR-98-5p mimic on the level of BACH1 (A) mRNA and (B) protein expression in human mesangial cells. (C) Effect of the BACH1-overexpression plasmid and co-transfection with the miR-98-5p mimic on cellular proliferation in human mesangial cells. (D) Effect of the BACH1-overexpression plasmid or co-transfection with the miR-98-5p mimic on the cellular secretion of TNF-alpha and IL-6 in human mesangial cells. All experiments were repeated three times and the data are expressed as the mean +- standard deviation. Data were analyzed by one-way ANOVA followed by a Tukey's post hoc test. ** P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 Transfection with the miR-98-5p inhibitor promotes BACH1 siRNA-induced human mesangial cell proliferation. Effect of BACH1 siRNA or co-transfection with the miR-98-5p inhibitor on (A) BACH1 mRNA and (B) protein expression in human mesangial cells. (C) Effect of BACH1 siRNA or co-transfection with the miR-98-5p mimic on human mesangial cell proliferation. (D) Effect of transfection with the BACH1 siRNA or co-transfection with the miR-98-5p inhibitor on the secretion of TNF-alpha and IL-6 of human mesangial cells. All experiments were repeated three times and the data are expressed as the mean +- standard deviation. Data were analyzed by one-way ANOVA followed by a Tukey's post hoc test. ** P

PA5-103060