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- Antigen structure
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- Validations
- Western blot [4]
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- Product number
- MA5-15646 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- ETV5 Monoclonal Antibody (3H3)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- MA5-15646 targets ETV5 in WB applications and shows reactivity with Human samples.
- Antibody clone number
- 3H3
Submitted references Chromatin accessibility analysis identifies the transcription factor ETV5 as a suppressor of adipose tissue macrophage activation in obesity.
Hu RD, Zhang W, Li L, Zuo ZQ, Ma M, Ma JF, Yin TT, Gao CY, Yang SH, Zhao ZB, Li ZJ, Qiao GB, Lian ZX, Qu K
Cell death & disease 2021 Oct 29;12(11):1023
Cell death & disease 2021 Oct 29;12(11):1023
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of ETV5 using ETV5 monoclonal antibody (Product # MA5-15646) in Jurkat (1), NIH/3T3 (2) and MCF-7 (3) cell lysate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of ETV5 using ETV5 monoclonal antibody (Product # MA5-15646) in Jurkat (1), NIH/3T3 (2) and MCF-7 (3) cell lysate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- KD of ETV5 was achieved by transfecting PC-3 with ETV5 specific siRNAs (Silencer® select Product # s4863, s4864). Western blot analysis (Fig. a) was performed using whole cell extracts from the ETV5 KD cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with ETV5 Monoclonal Antibody (3H3) (Product # MA5-15646, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to ETV5..
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-ETV5 Monoclonal Antibody (Product # MA5-15646) and a 57kDa band corresponding to ETV5 was observed across cell lines tested. An isoform corresponding to 62 kDa was observed only in PC-3 cell line. An uncharacterized band was observed at 100kDa in NTERA-2, HEK293, Daudi and HeLa. Whole cell extracts (30 µg lysate) of MCF-7 (Lane 1), NTERA-2 (Lane 2), SH-SY5Y (Lane 3), PC-3 (Lane 4), A549 (Lane 5), A-431 (Lane 6), HEK293 (Lane 7), Daudi (Lane 8) and HeLa (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence Goat Anti-Mouse IgG Secondary Antibody, HRP conjugate (Product # A28177, 1:4000 dilution)using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)..
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of ETV5 was performed using 70% confluent log phase A-431 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 ETV5 Monoclonal Antibody (3H3) (Product # MA5-15646) at 1:200 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) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominantly 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)
- Main image
- Experimental details
- Fig. 3 ETV5 expression was downregulated in ATMs from HFD-fed mice and in in vitro induced MMe. A Etv5 mRNA levels in ATMs from CD- ( n = 6) and HFD-fed ( n = 6) mice. B Etv5 mRNA levels in BMDMs after 24 h of stimulation with palmitate (P, 0.4 mM), glucose (G, 30 mM), and insulin (I, 10 nM). Gene expression levels were assessed by RT-qPCR. Data were pooled from two independent experiments. C Single-cell RNA-sequencing data of stromal vascular fraction from mouse iWAT. tSNE plot showing myeloid cell clusters. D Featureplot showing Lyz2 , Etv5 and Itgax expression by different myeloid cell clusters. E ETV5 protein levels and F quantification in Raw264.7 cells and BMDMs after treatment with P, G, I alone or in combination for 24 h. G ETV5 protein levels and H quantification in Raw264.7 cells and BMDMs after treatment with P, G, and I in pairs for 24 h. Data in E and G were representative of three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, Student's t test.
