- MA5-14801 - Invitrogen Antibodies SNAI1 antibody

Submitted references LINC00461 facilitates HNSCC development and reduces chemosensitivity by impairing miR-195-mediated inhibition of HOXA10.

Guan Y, Guan A, Chen L, Gong A
Molecular therapy oncolytics 2021 Jun 25;21:74-86

Loss of ZNF677 Expression Is an Independent Predictor for Distant Metastasis in Middle Eastern Papillary Thyroid Carcinoma Patients.

Siraj AK, Poyil PK, Parvathareddy SK, Alobaisi K, Ahmed SO, Al-Sobhi SS, Al-Dayel F, Al-Kuraya KS
International journal of molecular sciences 2021 Jul 22;22(15)

CHD4 Predicts Aggressiveness in PTC Patients and Promotes Cancer Stemness and EMT in PTC Cells.

Pratheeshkumar P, Siraj AK, Divya SP, Parvathareddy SK, Alobaisi K, Al-Sobhi SS, Al-Dayel F, Al-Kuraya KS
International journal of molecular sciences 2021 Jan 6;22(2)

Transmembrane protein ADAM29 facilitates cell proliferation, invasion and migration in clear cell renal cell carcinoma.

Li SL, Jiang TQ, Cao QW, Liu SM
Journal of chemotherapy (Florence, Italy) 2021 Feb;33(1):40-50

Diversity in the Extracellular Vesicle-Derived Microbiome of Tissues According to Tumor Progression in Pancreatic Cancer.

Jeong JY, Kim TB, Kim J, Choi HW, Kim EJ, Yoo HJ, Lee S, Jun HR, Yoo W, Kim S, Kim SC, Jun E
Cancers 2020 Aug 19;12(9)

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Snail in extracts from HTC116 and Hela cells using Snail monoclonal antibody (Product # MA5-14801).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of SNAIL was achieved by transfecting HCT 116 with SNAIL specific siRNAs (Silencer® select Product # s13186, s13187). Western blot analysis (Fig. a) was performed using whole cell extracts from the SNAIL knockdown cells (lane 2) and non-specific scrambled siRNA transfected cells (lane 1). The blots were probed with SNAIL Monoclonal Antibody (F.31.8) (Product # MA5-14801, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to SNAIL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on modified whole cell lysates (1% SDS) of A549 (Lane 1), A549 treated with TNFa (20ng/ml, 72hrs) (Lane 2), H-1975 (Lane 3) and HCT 116 (Lane 4). The blot was probed with SNAIL Monoclonal Antibody (F.31.8) (Product # MA5-14801, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25µg/ml, 1:4000 dilution). A 30 kDa band corresponding to SNAIL was induced upon TNFa treatment in A549 and was also observed in all other cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of SNAIL was performed using 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with SNAIL Monoclonal Antibody (F.31.8) (Product # MA5-14801) at 1:250 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 CHD4 activates epithelial-to-mesenchymal transition in PTC cells. ( A ) Forced expression of CHD4 activates EMT. K1 cells were transfected with either empty vector or CHD4 cDNA. After cell lysis, equal amounts of proteins were separated by SDS-PAGE, transferred to immobilon membrane, and immuno-blotted with antibodies against CHD4, E-cadherin, N-cadherin, Vimentin, Twist, Snail1, Zeb1, MMP-2, MMP-9 and GAPDH as indicated. ( B , C ) Forced expression of CHD4 increases invasion. K1 cells were transfected with either empty vector or CHD4 cDNA. Cells were seeded into the upper compartment of invasion chambers. The bottom chambers were filled with RPMI media. After 24 h incubation, invaded cells were fixed, stained and quantified. (Scale bars, 1 mm). ( D ) Forced expression of CHD4 increases migration. K1 cells were transfected with either empty vector or CHD4 cDNA. Cells were seeded into the upper compartment of migration chambers. The bottom chambers were filled with RPMI media. After 24 h incubation, migrated cells were fixed, stained and quantified. ( E ) Silencing of CHD4 down-regulates the expression of EMT markers in PTC cells. PTC cells were transfected with scrambled siRNA and CHD4 siRNA (50 and 100 nM). After 48 h, cells were lysed and proteins were immunoblotted with antibodies against CHD4, E-cadherin, N-cadherin, Vimentin, Twist, Snail1, Zeb1, MMP-2, MMP-9 and GAPDH. ( F ) Knockdown of CHD4 in expressing cells. ( G , H ) Knockdown of CHD4 decreases invasive a

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Methylation status of PTC cell lines by MSP for the E-cadherin gene. ( A ) Methylation status of three PTC cancer cell lines assessed by MSP for the E-cadherin gene. MSP analyses of both methylated (M) and unmethylated (U) reactions were amplified from bisulfite-treated DNA and run in a 2% agarose gel. ( B ) E-cadherin and Snail1 protein levels were determined by Western blotting in PTC cell lines. PTC cells were lysed and equal amounts of proteins were separated by SDS-PAGE, transferred to Immobilon membrane, and immunoblotted with antibodies against E-cadherin, Snail1 and GAPDH. ( C , D ) The demethylation restore E-cadherin expression in BCPAP and TPC-1 cells. PTC cells were treated with different doses (1, 2.5 and 5 muM) of 5-aza-2'deoxycytidine for 72 h and cells were lysed. Equal amounts of proteins were separated by SDS-PAGE, transferred to Immobilon membrane, and immunoblotted with antibodies against E-cadherin, Snail1 and GAPDH. ( E ) There was no change in the expression of E-cadherin and Snail1 in K1 cells after treatment with 5-aza-2'-deoxycytidine.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 Silencing of Snail1 and Zeb1 suppresses self-renewal ability of spheroids generated from PTC cells. ( A ) Knockdown of Snail1. PTC cells were transfected with Snail1 shRNA and knockdown was confirmed by immunoblotting. ( B , C ) Silencing of Snail1 suppresses spheroid growth. PTC cells were transfected with Snail1 shRNA and cells were subjected to sphere forming assay. Spheroids in the entire dish was counted. (Scale bars, 1 mm). ( D ) Silencing of Snail1 reduces stemness of spheroids. PTC cells were transfected with Snail1 shRNA and grown in sphere medium. Proteins were isolated from spheroids and immunoblotted with antibodies against Snail1, CD44, CD133, OCT4, NANOG and GAPDH. ( E ) Knockdown of Zeb1. PTC cells were transfected with Zeb1 shRNA and knockdown was confirmed by immunoblotting. ( F , G ) Silencing of Zeb1 suppresses spheroid growth. PTC cells were transfected with Zeb1 shRNA and cells were subjected to sphere forming assay. Spheroids in the entire dish was counted. (Scale bars, 1 mm). ( H ) Silencing of Zeb1 reduces stemness of spheroids. PTC cells were transfected with Zeb1 shRNA and grown in sphere medium. Proteins were isolated from spheroids and immunoblotted with antibodies against Zeb1, CD44, CD133, OCT4, NANOG and GAPDH. Data presented in the bar graphs are the mean +- SD of two independent experiments. * Indicates a statistically significant difference compared to control with p < 0.05.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 LINC00461 induces the EMT process in HNSCC (A) Immunofluorescence detection of EMT-related factors E-cadherin, Snail, and Vimentin in the HNSCC cell line FADU (400x). Three groups of red fluorescence represent E-cadherin, Snail, and Vimentin proteins, respectively, and blue fluorescence represents nuclei. (B) The protein bands of EMT-related factors E-cadherin, Snail, and Vimentin in the HNSCC cell line FADUs. (C) Quantitative analysis of (B). (D) The number of migrated FADU cells. (E) The number of invasive FADU cells. All of the data are measurement data and expressed as means +- standard deviations. The data among multiple groups were compared using 1-way ANOVA. The experiment was conducted 3 times independently. *p < 0.05 versus the pcDNA-NC group or the si-NC group.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 ZNF677 attenuates the metastatic potential of PTC cells. ( A - C ) Forced expression of ZNF677 decreased cell invasion and migration. ZNF677-overexpressing clones were seeded into the upper compartment of invasion or migration chambers. The bottom chambers were filled with RPMI media. After 24 h of incubation, invaded or migrated cells were fixed, stained, and quantified. ( D - F ) Demethylation of the ZNF677 gene decreased cell invasion and migration. PTC cells after treatment with the indicated dose of 5-aza-2'deoxycytidine for 72 h were used for invasion and migration experiments as described before. Data presented by the bar graphs are the mean +- SD of three independent experiments ( n = 3). * Indicates a statistically significant difference compared to control, with p < 0.05. ( G , H ) Forced expression of ZNF677 or demethylation of ZNF677 gene attenuates EMT in PTC cells. Protein extracts from ZNF677-overexpressing clones or cells after treatment with the indicated dose of 5-aza-2'deoxycytidine for 72 h were immuno-blotted with antibodies against ZNF677, E-cadherin, N-cadherin, Twist, Snail1, Zeb1, MMP-2, MMP-9, and GAPDH ( n = 3).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5. Effects of knockdown or overexpression of ADAM29 on proliferation- and motion-related proteins. Western blot assay was adopted to determine the effect of ADAM29 knockdown (a) or overexpression (b) on the level of Cyclin D1, PCNA, MMP9 and Snail. ** p < 0.01 vs. si-con group or vector group.

MA5-14801

Antibody data