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Ab200828

Manufactured by Abcam
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Ab200828 is a primary antibody product offered by Abcam. It is a recombinant monoclonal antibody targeting a specific protein. The antibody is produced in a mammalian cell line and is purified using protein A affinity chromatography.

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Lab products found in correlation

Ab207718, by Abcam (3 mentions) Ab15580, by Abcam (3 mentions) Ab92312, by Abcam (2 mentions) Ab110638, by Abcam (2 mentions) Ab92471, by Abcam (2 mentions) Ab109185, by Abcam (2 mentions) Ab212188, by Abcam (2 mentions) Ab3982, by Abcam (2 mentions) M01263 2, by Boster Bio (2 mentions) Protein a g plus agarose bead, by Santa Cruz Biotechnology (2 mentions) Igg antibody 5145s, by Cell Signaling Technology (2 mentions) Sc 23957, by Santa Cruz Biotechnology (2 mentions) Cytokeratin, by Biocare Medical (1 mentions) Ab210960, by Abcam (1 mentions) Ab227584, by Abcam (1 mentions) Ab182733, by Abcam (1 mentions) Ab182858, by Abcam (1 mentions) Anti rabbit igg as014, by ABclonal (1 mentions) Ab2302, by Abcam (1 mentions) Cl 0283, by Procell (1 mentions)

9 protocols using ab200828

1

Immunofluorescence Analysis of AGR2, WFDC2, and TSC22D3

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Immunofluorescence studies for the detection of AGR2 (1 in 100 dilution) (Abcam, Cat No: ab227584), WFDC2 (1 in 250 dilution) (Abcam, Cat No: ab200828) and TSC22D3 (1 in 50 dilution) (Abcam, Cat No: ab197987) were performed as described previously [24 (link),33 (link),34 (link)]. Epithelial expression of AGR2, WFDC2 and TSC22D3 positive cells was confirmed using cytokeratin (1 in 500) (Biocare, Denmark) and appropriate Alexa fluorophor conjugated secondary antibodies (Thermo Fisher Scientific, Austin, TX, USA). Localization of AGR2 protein expression to the endoplasmic reticulum (ER) was confirmed using the ER marker GRP94 (1 in 1000 dilution) (Abcam, Cat No: ab210960).
Alvarez X., Sestak K., Byrareddy S.N, & Mohan M. (2020). Long Term Delta-9-tetrahydrocannabinol Administration Inhibits Proinflammatory Responses in Minor Salivary Glands of Chronically Simian Immunodeficieny Virus Infected Rhesus Macaques. Viruses, 12(7), 713.
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2

Antibody Profiling for Cancer Markers

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The following antibodies were purchased from the respective suppliers: anti-RNPS1 (ab79233), anti-Ki-67 (ab15580), anti-CEA (ab207718), anti-CA199 (ab3982), anti-CA153 (ab109185), anti-HE4 (ab200828), anti-Bcl-2 (ab182858), anti-Bax (ab182733), anti-MLH1 (ab92312), anti-cleaved caspase-3 (ab2302), anti-MSH2 (ab212188), anti-MSH6 (ab92471), and anti-PMS2 (ab110638) (all from Abcam, Cambridge, USA) and anti-β-actin (M01263-2; Boster, Wuhan, China). The secondary antibodies used were anti-rabbit IgG (AS014) and anti-mouse IgG (H+L) (AS003), both from ABclonal (Wuhan, China), and IMR-1A (HY-100431A; MedChemExpress, Dallas, TX, USA).
Liu X., Ma H., Ma L., Li K, & Kang Y. (2021). RNA-binding protein with serine-rich domain 1 regulates microsatellite instability of uterine corpus endometrial adenocarcinoma. Clinics, 76, e3318.
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3

Endometrial Cell Line Characterization

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This study used normal endometrial cells (NECs), KLE cells, RL952 cells, Ishikawa cells, and ECC-1 cells. They were maintained and cultured as previously described [15 (link)]. The cell lines used in this study were as follows: normal endometrial cells (NEC, CP-H058, Procell, Wuhan, China), KLE (CL-0133, Procell, Wuhan, China), RL952 (CL-0197, Procell, Wuhan, China), Ishikawa (CL-0283, Procell, Wuhan, China), and ECC-1 (BS-C163325, BinSuiBio, Shanghai, China).
The following antibodies were used: Anti-METTL5 (16791-1-AP, Proteintech, Rosemont, USA), anti-Ki-67 (ab15580, Abcam, Cambridge, USA), anti-CEA (ab207718, Abcam, Cambridge, USA), anti-CA199 (ab3982, Abcam, Cambridge, USA), anti-CA153 (ab109185, Abcam, Cambridge, USA), anti-HE4 (13, ab200828, Abcam, Cambridge, USA), anti-MLH1 (ab92312, Abcam, Cambridge, USA), anti-MSH2 (ab212188, Abcam, Cambridge, USA), anti-MSH6 (ab92471, Abcam, Cambridge, USA), anti-PMS2 (ab110638, Abcam, Cambridge, USA), and anti-β-actin (M01263-2, Boster, Wuhan, China).
Liu X., Ma H., Ma L., Li K, & Kang Y. (2022). The potential role of methyltransferase-like 5 in deficient mismatch repair of uterine corpus endometrial carcinoma. Bioengineered, 13(3), 5525-5536.
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4

Multiplex Protein Expression in Lung Cancer

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The cell lines used were as follows: HAE (CP-H209, Procell, Wuhan, China), A549 (CL-0016, Procell), HCC827 (CL-0094, Procell), NCL-H1299 (CL-0165, Procell), and NCL-H524 (CL-0403, Procell).
The following antibodies were used: anti-TMED2 (ab251705, Abcam, Cambridge, USA), anti-Ki-67 (ab15580, Abcam), anti-CEA (ab207718, Abcam), anti-NSE (ab180943, Abcam), anti-EGFR (ab200828, Abcam), anti-TLR4 (ab13556, Abcam), anti-NF-κB-p-p65 (BM3940, Boster, Wuhan, China), anti-IL-1β (ab2105, Abcam); anti-IL-18 (ab207323, Abcam); and anti-β-actin (M01263-2, Boster). The secondary antibodies used were anti-rabbit IgG (AS014, ABclonal, Wuhan, China) and anti-mouse IgG (H+L) (AS003, ABclonal).
Feng L., Cheng P., Feng Z, & Zhang X. (2022). Transmembrane p24 trafficking protein 2 regulates inflammation through the TLR4/NF-κB signaling pathway in lung adenocarcinoma. World Journal of Surgical Oncology, 20, 32.
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5

Immunoprecipitation of HE4 Protein

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A total of 2 μg of anti-HE4 antibody (abcam: ab200828) or anti-rabbit IgG antibody (Zhongshan Jinqiao) was added to 500 μg protein lysate and incubated at 4 °C overnight. The next day, 40 μl protein A/G PLUS-Agarose beads (Santa Cruz: sc-2003) was added at 4 °C. After 6 h, the immunoprecipitated protein complex was added with 2 × loading buffer and boiled to denature the protein and separate it from the protein-G beads. The same method was performed using 2 μg of the anti-ZNF703 primary antibody to precipitate HE4, and the precipitated protein was detected using western blotting with the anti-HE4 primary antibody.
Wang S., Wang C., Hu Y., Li X., Jin S., Liu O., Gou R., Zhuang Y., Guo Q., Nie X., Zhu L., Liu J, & Lin B. (2020). ZNF703 promotes tumor progression in ovarian cancer by interacting with HE4 and epigenetically regulating PEA15. Journal of Experimental & Clinical Cancer Research : CR, 39, 264.
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6

Immunoprecipitation and Protein Extraction

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Cells in the exponential growth phase were collected and washed. Ice-cold lysis buffer was added, and the cell suspension was sonicated. The supernatant was collected and the total protein concentration was determined. Samples (500 μg) were then incubated with 2 μg of YWHAE (1:100, sc-23957, Santa Cruz Biotechnology, Santa Cruz, CA) or HE4 (1:1500, ab200828, Abcam, Cambridge, UK) primary antibodies. An IgG antibody (5145S, Cell Signaling Technology Inc., Danvers, MA) of the same species as the primary antibodies was used as a negative control. Following mixing, the samples were rotated slowly overnight at 4 °C. Subsequently, Protein A/G Plus-Agarose Beads (sc-2003, Santa Cruz Biotechnology, Santa Cruz, CA) were added to each tube and incubated for 6 h. Bound proteins were collected via centrifugation, (2×) loading buffer was added, and the samples were boiled to denature the proteins.
Li X., Wang C., Wang S., Hu Y., Jin S., Liu O., Gou R., Nie X., Liu J, & Lin B. (2021). YWHAE as an HE4 interacting protein can influence the malignant behaviour of ovarian cancer by regulating the PI3K/AKT and MAPK pathways. Cancer Cell International, 21, 302.
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7

Immunohistochemical Staining of HE4 in Lung

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IHC staining was performed in lung specimens fixed with 10% formalin and embedded in paraffin before being cut into 5-µm-thick sections. After dewaxing and rehydration, the sections were incubated with 3% H2O2 for 5 minutes to block the activity of peroxidase. The sections were then incubated with rabbit monoclonal HE4 antibody (ab200828; Abcam, Tokyo, Japan) at 4 ℃ overnight, followed by secondary antibody (Vector Laboratories, Burlingame, CA, USA) at 25 ℃ for 60 minutes. Subsequently, the visualization of immunoreaction was conducted by 3,3-diaminobenzidine (DAB) chromogen solution (DAB substrate kit; Vector Laboratories) and then counterstained with hematoxylin.
Tian M., Meng K., Gao Y., Zhang J., Xie M., Tian Y., Liu X., Ma M., Cai Y., Wu H., Ding J., Chen J, & Cai H. (2022). Elevated serum human epididymis protein 4 is associated with disease severity and worse survival in idiopathic pulmonary fibrosis: a cohort study. Annals of Translational Medicine, 10(18), 992.
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8

Prostate Cancer Cell Line Characterization

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Human prostate cancer PC-3 and DU-145 cell lines were cultured in RPMI-1640 with 10% fetal bovine serum (FBS). The 293 T cell line was cultured in DMEM with 10% FBS. PC-3, DU-145, and 293 T cell lines were obtained from the American Type Culture Collection and were recently authenticated.
Antibodies against FLAG (F1804, Sigma), HA (TA180128, OriGene), GAPDH (sc-365062, Santa Cruz), WFDC2 (ab200828, Abcam), EGFR (ab52894, Abcam), p-EGFR (4407 S, CST), AKT (4691 L, CST), p-AKT (4060 L, CST), c-Myc (ab32072, Abcam), E2F-1 (ab179445, Abcam), GSK3B (12456 S, CST), p-GSK3B (5558 S, CST), Snail (3879 S, CST), E-cad (3195 S, CST), N-cad (13116 S, CST), Vimentin (5741 S, CST), and secreted protein HE4 (CSB-DP018B, CUSABIO), EGF (PHG0311L, Gibco) were purchased from indicated commercial sources.
Xiong Y., Yuan L., Chen S., Xu H., Peng T., Ju L., Wang G., Xiao Y, & Wang X. (2020). WFDC2 suppresses prostate cancer metastasis by modulating EGFR signaling inactivation. Cell Death & Disease, 11(7), 537.
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9

Immunoprecipitation of YWHAE and HE4

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Cells in exponential growth were collected and washed. Iced lysis buffer was added and the cell suspension was sonicated. The supernatant was collected and total protein concentration was determined. Total protein samples (500 μg) were incubated with 2 μg of YWHAE (1 100, sc-23957, Santa Cruz Biotechnology, Santa Cruz, CA) or HE4 (1:1500, ab200828, Abcam, Cambridge, UK) primary antibody. An IgG antibody (5145S, Cell Signaling Technology, Beverly, MA) of the same species of the primary antibody was used as negative control. After mixing, the samples were left overnight at 4°C with slow rotation. Afterwards, beads (sc-2003, Santa Cruz Biotechnology, Santa Cruz, CA) were added to each tube and incubated for 6 h. The bound proteins were collected by centrifugation, (2×) Loading Buffer was added, and the samples were heated to denature the proteins.
Li X., Wang C., Wang S., Hu Y., Jin S., Liu O., Gou R., Nie X., Liu J., & Lin B. (2021). YWHAE Influences the Malignant Behaviour of Ovarian Cancer by Regulating the PI3K/AKT and MAPK Pathways Via HE4.
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