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Ab102010

Manufactured by Abcam
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Ab102010 is a high-quality primary antibody produced by Abcam. It is designed for use in various immunoassay applications, such as ELISA, Western blotting, and immunohistochemistry. The antibody is specific for a target protein, and its core function is to detect and bind to that protein in biological samples.

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Lab products found in correlation

A11008, by Thermo Fisher Scientific (2 mentions) A 11030, by Thermo Fisher Scientific (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) A11081, by Thermo Fisher Scientific (2 mentions) Spe100, by Leica (2 mentions) Ab6640, by Abcam (2 mentions) A 11056, by Thermo Fisher Scientific (2 mentions) Ab82527, by Abcam (2 mentions) Ab84593, by Abcam (2 mentions) Sc 30820, by Santa Cruz Biotechnology (2 mentions) Ab46154, by Abcam (2 mentions) Ideal chip seq kit for transcription factors, by Diagenode (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Ab76003, by Abcam (1 mentions) Ab92536, by Abcam (1 mentions) Bs 10297r, by Bioss Antibodies (1 mentions) Ab9669, by Abcam (1 mentions) Ab9669, by Santa Cruz Biotechnology (1 mentions) Ecl solution, by Cytiva (1 mentions) Ab81086, by Abcam (1 mentions)

9 protocols using ab102010

1

Genotyping and Protein Expression of Etv5 in Mice

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For CRISPR-induced mutation assays, female and male heterozygous edited mice were bred to generate homozygous edited offspring. DNA was extracted from the toes of pups from the same litter at day 2 post-birth for PCR analysis and sequencing. The following two primers were used for genotyping by PCR: mouse Etv5 primer-Forward: 5′-CAACTGGTGCCCTTCCCAGTCT-3′, mouse Etv5 primer-Reverse: 5′-GCCGCTCTTAAACCTGTTCATTCG-3′. WT mice were designated as the control group, and Etv5−/− mice comprised the experimental group. Polyclonal anti-ERM/Etv5 antibody (1:100 dilution; ab102010, Abcam, Cambridge, MA, USA) recognizing amino acids 244 of the human ERM/Etv5 was used to measure Etv5 protein expression in Etv5−/− male and WT mice by western blotting. The anti-β-actin monoclonal antibody was used as the loading control.
Zhang X., Zhao X., Li G., Zhang M., Xing P., Li Z., Chen B., Yang H, & Wu Z. (2021). Establishment of Etv5 gene knockout mice as a recipient model for spermatogonial stem cell transplantation. Biology Open, 10(1), bio056804.
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2

ETV5 Chromatin Immunoprecipitation Protocol

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ETV5 Chromatin ImmunoPrecipitation (ChIP) was performed using the iDeal ChIP-seq kit for Transcription Factors (Diagenode) and the anti-ETV5 rabbit polyclonal antibody ab102010 from Abcam, using the same procedure as previously described [47 (link)]. The TruSeq ChIP library preparation kit was used to prepare Illumina sequencing libraries from the ChIP and input DNAs, as described [47 (link)].
Lopez-Delisle L., Pierre-Eugène C., Louis-Brennetot C., Surdez D., Raynal V., Baulande S., Boeva V., Grossetête-Lalami S., Combaret V., Peuchmaur M., Delattre O, & Janoueix-Lerosey I. (2018). Activated ALK signals through the ERK–ETV5–RET pathway to drive neuroblastoma oncogenesis. Oncogene, 37(11), 1417-1429.
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3

Protein Expression Analysis of Cell Lines

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After being washed by PBS, all kinds of cell lines were harvested in RIPA buffer. The protein concentrations were measured using Bradford assay. Samples were separated on 8-12% SDS-PAGE gels and then transferred to PVDF membrane (Millipore, USA) and probed with primary antibodies specific for ETV5 (ab102010, Abcam), MMP2 (ab92536, Abcam), MMP9 (ab76003, Abcam), SKA1 (bs-7846R, Bioss) and TRPV2 (bs-10297R, Bioss). β-Actin, Tubulin and GAPDH were used as loading controls for western blots.
Sun M.C., Fang K., Li Z.X., Chu Y., Xu A.P., Zhao Z.Y., Leng Z.Y., Zhang Y.W., Zhang Z.H., Zhang L., Chen T, & Xu M.D. (2022). ETV5 overexpression promotes progression of esophageal squamous cell carcinoma by upregulating SKA1 and TRPV2. International Journal of Medical Sciences, 19(6), 1072-1081.
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4

Histological and Immunohistochemical Analysis of Testis and Epididymis

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Tissues were harvested at necropsy and fixed overnight in 4% paraformaldehyde (PFA) in PBS. Subsequently, 5 μm-thick testis and epididymal sections from infected and uninfected mice were processed for histology by H & E staining. For immunohistochemistry, the tissue sections were incubated with mouse primary monoclonal anti-CD45 (610266; BD Biosciences), anti-ETV5 (ab102010; Abcam), and anti-GATA4 (ab84593; Abcam), rabbit polyclonal anti-Lin28a (3978S, Cell Signaling), rat polyclonal anti-TRA98 (ab82527; Abcam), rat polyclonal anti-F4/80 (ab6640; Abcam), or goat polyclonal anti-3β-HSD antibodies (SC-30820, Santa Cruz Biotechnology). After washing, slides were stained with Alexa Fluor 488- or, Alexa Fluor 546-conjugated goat anti-rabbit, goat anti-mouse, or donkey anti-goat (1:1000; A11008, A11081, A11030, or A11056; ThermoFisher Scientific) secondary antibodies for 1 h, and mounted with prolong gold anti-fade mount containing the nuclear counter stain, DAPI (ThermoFisher Scientific). Immunostaining was detected by confocal microscopy (Leica SPE100, Germany).
Govero J., Esakky P., Scheaffer S.M., Fernandez E., Drury A., Platt D.J., Gorman M.J., Richner J.M., Caine E.A., Salazar V., Moley K.H, & Diamond M.S. (2016). Zika virus infection damages the testes in mice. Nature, 540(7633), 438-442.
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5

Immunohistochemical Analysis of ETV5 in ESCC

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This study was approved by the institutional review board of Shanghai East Hospital, Tongji University. ESCC and corresponding healthy esophageal mucosa (CHEM) tissues of 79 patients were collected. No patient received previous treatment, such as surgery, chemotherapy, and radiotherapy. The details of Clinicopathological factors were shown in supplementary table 2. After being fixed by Formalin and embedded by paraffin, tissue sections were cut to 4μm thickness and then placed in xylene and graded alcohols for deparaffinization and hydration. We performed heat-induced antigen retrieval in EDTA (PH 8.0) buffer for 15 minutes by using a microwave oven. To reduce nonspecific staining, we performed blocking with 10% goat serum. After the specific primary anti-ETV5 (ab102010, Abcam) was dropped onto the sections and incubated overnight at 4℃, the slides were counterstained with light hematoxylin, dehydrated, and cover-slipped.
Sun M.C., Fang K., Li Z.X., Chu Y., Xu A.P., Zhao Z.Y., Leng Z.Y., Zhang Y.W., Zhang Z.H., Zhang L., Chen T, & Xu M.D. (2022). ETV5 overexpression promotes progression of esophageal squamous cell carcinoma by upregulating SKA1 and TRPV2. International Journal of Medical Sciences, 19(6), 1072-1081.
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6

Histological and Immunohistochemical Analysis of Testis and Epididymis

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Tissues were harvested at necropsy and fixed overnight in 4% paraformaldehyde (PFA) in PBS. Subsequently, 5 μm-thick testis and epididymal sections from infected and uninfected mice were processed for histology by H & E staining. For immunohistochemistry, the tissue sections were incubated with mouse primary monoclonal anti-CD45 (610266; BD Biosciences), anti-ETV5 (ab102010; Abcam), and anti-GATA4 (ab84593; Abcam), rabbit polyclonal anti-Lin28a (3978S, Cell Signaling), rat polyclonal anti-TRA98 (ab82527; Abcam), rat polyclonal anti-F4/80 (ab6640; Abcam), or goat polyclonal anti-3β-HSD antibodies (SC-30820, Santa Cruz Biotechnology). After washing, slides were stained with Alexa Fluor 488- or, Alexa Fluor 546-conjugated goat anti-rabbit, goat anti-mouse, or donkey anti-goat (1:1000; A11008, A11081, A11030, or A11056; ThermoFisher Scientific) secondary antibodies for 1 h, and mounted with prolong gold anti-fade mount containing the nuclear counter stain, DAPI (ThermoFisher Scientific). Immunostaining was detected by confocal microscopy (Leica SPE100, Germany).
Govero J., Esakky P., Scheaffer S.M., Fernandez E., Drury A., Platt D.J., Gorman M.J., Richner J.M., Caine E.A., Salazar V., Moley K.H, & Diamond M.S. (2016). Zika virus infection damages the testes in mice. Nature, 540(7633), 438-442.
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7

Immunohistochemical Analysis of Xenograft Tumors

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Immunohistochemistry analysis of xenograft tumors in nude mice and CRC specimens were conducted as previously described3 (link),19 (link). The sections were incubated with antibodies against ETV5 (ab102010, Abcam, Cambridge, UK), VEGFA (ab46154, Abcam, Cambridge, UK), CD31 (3528, CST, 1:100, Boston, USA), Ki67 (1:200, Santa Cruz, Dallas, USA), and CCL2 (ab9669, Abcam, Cambridge, UK).
Feng H., Liu K., Shen X., Liang J., Wang C., Qiu W., Cheng X, & Zhao R. (2020). Targeting tumor cell-derived CCL2 as a strategy to overcome Bevacizumab resistance in ETV5+ colorectal cancer. Cell Death & Disease, 11(10), 916.
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8

Immunohistochemical Profiling of Tumor Samples

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FFPE slides with CRC specimens and nude mice tumor tissue sections were stained as previously described [3, 19] . Antibodies used for IHC included antibodies against ETV5 (ab102010, Abcam), VEGFA (ab46154, Abcam), CD31 (3528, Cell Signaling Technology), Ki67 (1:200, Santa Cruz), and CCL2 (ab9669).
Feng H., Liu K., Liang J., Wang C., Qiu W., Cheng X., & Zhao R. (2020). Targeting tumor cell-derived CCL2 as a strategy to overcome bevacizumab resistance in ETV5+ colorectal cancer.
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9

Protein Detection by Western Blotting

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Tissue samples were homogenized, and proteins were detected using Western blotting [24] .
Briefly, total proteins was separated by SDS-PAGE and transferred to the nitrocellulose membrane. Blots were blocked and then incubated with primer antibodies including anti-actin
(1:10000, Abcam A2066), anti-PEA3 (1:500, Abcam, ab101455), anti-ERM (1:1000, Abcam, ab102010) and anti-ER81 (1:1000, Abcam, ab81086). After washing, HRP conjugated secondary antibodies were applied for 1 hr at room temperature before applying ECL solution (Amersham Biosciences), exposed to film for 10-20 min, and developed by hand. Protein bands were quantified using GeneSnap acquisition and GeneTools analysis software.
Kandemir B., Davis S., Yiğit E.N., Öztürk G., Yılmaz B., Laroche S, & Kurnaz I.A. (2020). Expression of Pea3 protein subfamily members in hippocampus and potential regulation following neuronal stimulation. Neuroscience letters, 738.
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