Anti-MTA1 Antibody Picoband®

Western blot analysis of MTA1 using anti-MTA1 antibody (PA1483).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human MCF-7 whole cell lysates,
Lane 2: human HEK293 whole cell lysates,
Lane 3: human HELA whole cell lysates,
Lane 4: human PC-3 whole cell lysates,
Lane 5: human Raji whole cell lysates,
Lane 6: human K562 whole cell lysates,
Lane 7: rat brain tissue lysates,
Lane 8: mouse brain tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-MTA1 antigen affinity purified polyclonal antibody (Catalog # PA1483) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for MTA1 at approximately 80KD. The expected band size for MTA1 is at 80KD.

The expression of MTA1 is up-regulated in RCCs. a Immunohistochemistry was used to characterize the expression of MTA1 in RCC and adjacent tissues. MTA1 was highly expressed in ccRCCs, compared to very weak staining in adjacent tissue. b Image Pro Plus was used for the statistical analysis of the positive signal (IOD) of MTA1 expression in ccRCCs and the adjacent tissue. The expression of MTA1 was significantly up-regulated in ccRCCs. ** p < 0.01; scale bar: 100 µm and 50 µm. c The expression of MTA1 in RCC cell lines. Normal renal cell line HEK293T and RCC cell lines A498 and 768-O cell lysates were used in western blotting analysis with MTA1 and GAPDH antibodies. The bands of MTA1 were from Additional file : and gapdh were from Additional file :
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MTA1 promotes A498 cell migration. A498 cells were transfected with pcDNA3.1-Flag, Flag-MTA1, siNC, and si-MTA1. After 24 h, cells in the four conditions were subjected to the wound healing assay. Macrographs were taken under ×100 magnification
Index in PubMed under a CC BY license. PMID: 33059651

MTA1 enhances the invasion of A498 cells. a A498 cells were transfected with pcDNA3.1-Flag, Flag-MTA1, siNC, and si-MTA1. After 24 h, the transwell cell invasion assay using A498 cells was performed, and macrographs were taken under ×40 magnification. Scale bar: 50 µm. b Image J was used to count the transmigrated cells and Student’s t test to analyze differences. ** p < 0.01; *** p < 0.001
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MTA1 regulated the expression of metastasis-related factors via the NFκB pathway. A498 cells were transfected with pcDNA3.1-Flag, Flag-MTA1, siNC, and si-MTA1. a Protein levels of MTA1; cells were collected for WB with MTA1 and GAPDH antibodies. The bands of over-expressed MTA1 were from Additional file : Fig. 2A and down-expressed MTA1 were from Additional file : Fig. 2B with arrow. b Cells were lysed and used in qRT-PCR assays to measure the E-cadherin mRNA expression. Statistical analysis was performed using Student’s t test. b , c A498 cells were transfected with pcDNA3.1-Flag and Flag-MTA1. After 36 h, the Flag-MTA1 group was treated with 10 nM PDTC for 12 h. Then cells were harvested for qRT-PCR evaluation to measure the mRNA expression of MMP2 ( b ) and MMP9 ( c ). Statistical analysis was performed using ANOVA. d A498 cells were transfected with pcDNA3.1-Flag, Flag-MTA1, siNC, and si-MTA1. After 48 h, cells were subjected to western blotting analysis using antibodies targeting p-p65, p65 and ACTB. The bands of p-p65, p65 and ACTB were from Additional file : Fig. 3. e, f Image J was used to calculate the gray scanned bands of p-p65 ( b ) and p65 (C). * p < 0.05; ** p < 0.01; *** p < 0.001, **** p < 0.0001; NS not significant
Index in PubMed under a CC BY license. PMID: 33059651

MTA1 regulated the migration and invasion of RCC cells via NF-κB. A498 cells were transfected with pcDNA3.1-Flag, Flag-MTA1, and Flag-MTA1 + PDTC (10 nM). a In the wound healing assay, MTA1 improved the migration of RCC cells, while the addition of the inhibitor PDTC blocked the effect of MTA1 on migration. Scale bar: 100×. b In the transwell assay, MTA1 markedly induced invasion of A498 cells, but compared to the MTA1 group, the invasion of MTA1 + PDTC A498 cells was lower. ** p < 0.01; *** p < 0.001; Scale bar: ×40
Index in PubMed under a CC BY license. PMID: 33059651

Anti-MTA1 antibody, PA1483, IHC(P)
IHC(P): Human Rectal Cancer Tissue

IHC analysis of MTA1 using anti-MTA1 antibody (PA1483).
MTA1 was detected in paraffin-embedded section of rat ovary tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-MTA1Antibody (PA1483) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Anti-MTA1 antibody, PA1483, IHC(F)
IHC(F): Rat Ovary Tissue

IHC analysis of MTA1 using anti-MTA1 antibody (PA1483).
MTA1 was detected in paraffin-embedded section of rat ovary tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-MTA1 Antibody (PA1483) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

IHC analysis of MTA1 using anti-MTA1 antibody (PA1483).
MTA1 was detected in frozen section of mouse brain tissues. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-MTA1 Antibody (PA1483) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

IHC analysis of MTA1 using anti-MTA1 antibody (PA1483).
MTA1 was detected in paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-MTA1 Antibody (PA1483) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

IHC analysis of MTA1 using anti-MTA1 antibody (PA1483).
MTA1 was detected in paraffin-embedded section of human ovarian cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-MTA1 Antibody (PA1483) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

IHC analysis of MTA1 using anti-MTA1 antibody (PA1483).
MTA1 was detected in paraffin-embedded section of human pancrease cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-MTA1 Antibody (PA1483) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.