MA5-14154

antibody from Invitrogen Antibodies

Targeting: MITF bHLHe32, MI, WS2, WS2A

Provider product page for MA5-14154

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [40]
Comments [0]
Validations
Immunohistochemistry [1]
Other assay [3]

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Product number: MA5-14154 - Provider product page
Provider: Invitrogen Antibodies
Product name: MiTF Monoclonal Antibody (D5)
Antibody type: Monoclonal
Antigen: Other
Description: MA5-14154 targets Microphthalmia-Transcription Factor in GS and IHC (P) applications and shows reactivity with Canine and Human samples.
Antibody clone number: D5
Concentration: 0.2 mg/mL

MITF protein structure - MA5-14154 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1. MITF is highly expressed in kidney angiomyolipomas. (A) RNA-seq data showing MITF expression in 28 kidney angiomyolipomas compared to 2463 tumors of 27 different histologic types (TCGA dataset). Abbreviations for all TCGA tumor types are available here: https://gdc.cancer.gov/resources-tcga-users/tcga-code-tables/tcga-study-abbreviations . Gene expression is shown in RSEM values. (B) RNA-seq data showing MITF expression in 28 kidney angiomyolipoma compared to GTEx human normal tissues (~8,500 samples from 30 normal tissue types, v6p release). Gene expression is shown in FPKM values. (C) Representative IHC images show that MITF is expressed weakly if at all in normal kidney and highly expressed with nuclear localization in kidney angiomyolipoma. 7 kidney angiomyolipomas were examined and 6 of 7 (86%) showed MITF expression. Scale bar, 50 mum. (D) Immunoblot analysis of MITF in total protein lysates from 3 normal kidneys and 3 kidney angiomyolipoma tumors. (E) H3K27ac ChIP-seq data for 3 kidney angiomyolipomas, the human melanoma SK-MEL30 cell line, and one normal kidney sample for the MITF locus viewed in IGV. Peaks reflecting read density for H3K27ac are seen in all three angiomyolipomas near the TSS of MITF-A , while few reads are seen in this region in the melanoma cell line and normal kidney. The Super-enhancer region, highly marked for H3K27ac and identified by ROSE analysis near the MITF-A TSS is shown at higher magnification at bottom (IGV 10kb window). Two of

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2. MITF regulates cell growth, proliferation, migration and invasion in 621-101 cells. (A) Q-RT-PCR analysis of MITF expression after MITF knockout by CRISPR/Cas9 ( MITF.KO ) or overexpression of MITF ( MITF.OE ) compared with control (EV) in 621.101. beta-actin was used as a control. Each bar represents the mean +- SEM ( n = 4 per group; * p < 0.05; ** p < 0.01; *** p < 0.001). Data for 4 separate clones is shown. (B) Immunoblot analysis of 621-101 cells in which MITF has been knocked out by CRISPR/Cas9 (KO). Beta-actin serves as a loading control (3 independent experiments performed). (C) Immunoblot analysis of 621-101 cells in which MITF is stably overexpressed. beta-actin serves as a loading control (3 independent experiments performed). Data for 4 separate clones is shown. (D) Cell proliferation (Trypan blue assays) of 621-101 after MITF knockout or overexpression compared with control (EV) for indicated time points (hours). Each data point represents the mean of 3 independent experiments +- standard error of the mean (SEM) (** p < 0.01; *** p < 0.001). (E) Q-RT-PCR analysis of MITF expression after MITF silencing by 3 different siRNA oligos (si. MITF ) compared with control (si.CTRL) in 621.101 cells. Error bars represent +- SEM of triplicate wells from a representative experiment (** p < 0.01; *** p < 0.001). (F) Immunoblot analysis of MITF expression after MITF silencing (3 independent siRNA), along with siRNA controls in 621-101 cells. beta-actin serves as a l

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 10 The NGFR High state is associated with resistance to MAPK inhibitors in a subset of melanoma patients Immunohistochemical analysis of vemurafenib-naive tumors from three melanoma patients stained for NGFR, MITF, and Ki-67 (see Materials and Methods for patient clinical information). Covariate single-cell analysis of Ki-67 versus NGFR measured by immunofluorescence in pre-treatment, on-treatment (with dabrafenib and trametinib combination for 2 weeks), and post-relapse tumor biopsies of a BRAF -mutant melanoma patient (see Materials and Methods for patient clinical information). Cell population histograms representing c-Jun variations measured by immunofluorescence in the same patient-matched biopsies as shown in (B). NGFR gene expression changes in 21 matched pairs of pre-treatment and post-resistance tumor biopsies analyzed by RNA sequencing. MITF changes are shown for tumors with a post-resistance NGFR increase (increase = log 2 (fold-change) > 0.5, decrease = log 2 (fold-change) < -0.5, no change = |log 2 (fold-change)|