Mouse anti ha

Manufactured by Abmart

Sourced in China

Mouse anti-HA is a monoclonal antibody that binds specifically to the Hemagglutinin (HA) tag, a commonly used protein tag. It is designed for the detection and purification of HA-tagged recombinant proteins.

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Lab products found in correlation

Close spelling variants of this product

Anti-HA (31 citations) Mouse anti-HA mAb (4 citations) Mouse anti-Flag/HA-tags beads (2 citations)

6 protocols using mouse anti ha

Western Blot Analysis of Protein Expression

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Cells were lysed through cell lysis buffer with phenylmethylsulfonyl fluoride (PMSF; Beyotime, China). The samples were performed by SDS-PAGE and western blot. Briefly, the cell lysates were resolved in 12% polyacrylamide gel, and then transferred to PVDF membranes (Millipore, China). After blocked in Tris-buffered saline containing 0.1% Tween-20 (TBST) and 10% nonfat milk for 2 h, the membranes were incubated at 4°C for 12 h with the appropriate primary antibody (mouse anti-Flag (Sigma), rabbit anti-myc (Sigma), mouse anti-HA (Abmart, China), rabbit anti-GAPDH (Abmart), mouse anti-IRF3 (CST, China), or mouse anti-spho-IRF3 (CST)). Membranes were washed twice with TBST and incubated with HRP-labeled goat anti-rabbit or goat anti-mouse secondary antibody (Abmart) at room temperature for 1 h, and then visualized with Tanon 5200 chemiluminescence imaging system.

Mou C., Pan S., Wu H, & Chen Z. (2021). Disruption of interferon-β production by the Npro of atypical porcine pestivirus. Virulence, 12(1), 654-665.

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Antibody Characterization Protocol

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The following commercial antibodies were used: rabbit anti-Vps34 (PTG), rabbit anti-FBXL20 (Abcam), goat anti-FBXL20 (Santa Cruz Biotechnology), rabbit anti-p53 (PTG), rabbit anti-LC3 (Sigma), mouse anti-Beclin1 (Santa Cruz Biotechnology), rabbit anti-Atg14 (MBL), rabbit anti-GFP (Santa Cruz Biotechnology), mouse anti-GFP (Abmart), mouse anti-HA (Abmart), rabbit anti-Atg12–Atg5 (Epitomics), rabbit anti-CUL1 (Epitomics), rabbit anti-Skp1 (Epitomics), mouse anti-Flag (Sigma), mouse anti-Myc (Sigma), rabbit anti-ubiquitin (Dako), and rabbit anti-Tubulin (MBL). Chemicals were from Sigma.

Xiao J., Zhang T., Xu D., Wang H., Cai Y., Jin T., Liu M., Jin M., Wu K, & Yuan J. (2015). FBXL20-mediated Vps34 ubiquitination as a p53 controlled checkpoint in regulating autophagy and receptor degradation. Genes & Development, 29(2), 184-196.

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Immunostaining of Drosophila Tissues

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Fixation and antibody staining in imaginal discs were performed as described [70 (link)]. Fixation and antibody staining in cultured cells were performed as described [71 (link)]. Fixation and antibody staining in midguts were performed as described [37 (link)]. Primary antibodies used for the immunostaining were: mouse anti-Wdp (1:1000), chicken anti-lacZ (Abcam, 1:1000), mouse anti-Dl (DSHB, 1:50), mouse anti-Pros (DSHB, 1:200), rabbit anti-PH3 (Millipore, 1: 2000), mouse anti Brdu (DSHB, 1:200), rabbit anti Pdm1(1:1000, gift from Xiaohang Yang), mouse anti-V5 (Invitrogen, 1:3000), mouse anti-HA (Abmart, 1:500), rabbit anti-GM130 (Abcam, 1:200), rabbit anti-Rab5 (Abcam, 1:200), Gp anti-Sens (1:200), rabbit anti-Sal (1:100), and Rat anti-Ci (DSHB, 1:5). The primary antibodies were detected by fluorescent-conjugated secondary antibodies from Jackson ImmunoResearch Laboratories, Inc. The primary antibodies used for IP and western blot were: rabbit anti-V5 (Sigma, 1:1000), rabbit anti-HA (Santa Cruz, 1:1000), mouse anti-Wdp (1:500), rabbit anti-GFP (Abmart, 1:1000) and mouse anti-tubulin (Abmart, 1:1000).

Ren W., Zhang Y., Li M., Wu L., Wang G., Baeg G.H., You J., Li Z, & Lin X. (2015). Windpipe Controls Drosophila Intestinal Homeostasis by Regulating JAK/STAT Pathway via Promoting Receptor Endocytosis and Lysosomal Degradation. PLoS Genetics, 11(4), e1005180.

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Antibody-based Detection of Viral Proteins

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The following primary antibodies were used: PRRSV N protein antibody (JNT, China; no. JN0401), mouse anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH) antibody (TransGen Biotech, China; no. HC301), rabbit anti-HA (Cell Signaling Technology, USA; no. 3724 T), rabbit anti-Flag (Abmart, China; no. T20008M), mouse anti-HA (Abmart; no. M20003M), mouse anti-Flag (Beyotime, China; no. AF519), rabbit anti-GFP (Proteintech; no. 50430-2-AP), mouse anti-GFP (Proteintech; no. 66002-Ig), rabbit anti-β-actin (Proteintech; no. 81115-1-RR) and mouse anti-dsRNA (Scicons, Hungary; no. 10010500). The anti-ZNF283 antibody was generated by immunizing rabbits with bacterially expressed full-length porcine ZNF283.

Yi H., Ye R., Xie E., Lu L., Wang Q., Wang S., Sun Y., Tian T., Qiu Y., Wu Q., Zhang G, & Wang H. (2024). ZNF283, a Krüppel-associated box zinc finger protein, inhibits RNA synthesis of porcine reproductive and respiratory syndrome virus by interacting with Nsp9 and Nsp10. Veterinary Research, 55, 9.

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Validating Proteomics Findings with Western Blot

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To validate the proteomics results, a Western blot was conducted. Firstly, the proteins from each group were extracted and separated using 12% SDS-PAGE and transferred to a polyvinylidene fluoride (PVDF) membrane using a semi-dry transfer cell. The membrane was rinsed consecutively in Tris buffered saline (TBS) and 5% skimmed milk in TBS containing 0.5% Tween 20 (TBST) for 2 h at room temperature with 50 rpm shaking, followed by incubated with 1:2500 diluted mouse anti-HA (Catalog number:26D11, Abmart, Shanghai, China) as the primary antibody at room temperature for 3 h. After being washed three times with TBST, the membranes were incubated with goat-anti mouse IgG (1:2500) as the secondary antibody at room temperature for 1 h. After washing three times with TBST, the color development was performed using enhanced chemiluminescence (ECL) substrate (Solarbio, Beijing, China) in accordance with the manufacturer’s instructions. Finally, the PVDF membranes were stained with Commassie blue (CBB) to verify that the loading amounts were equal.

Xu L., Liu H., Zhu S., Meng Y., Wang Y., Li J., Zhang F, & Huang L. (2023). VmPacC-mediated pH regulation of Valsa mali confers to host acidification identified by comparative proteomics analysis. Stress Biology, 3(1), 18.

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Yeast Protein Extraction and Western Blotting

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Yeast intracellular proteins were extracted using a yeast total protein extraction kit (Bestbio, Shanghai, China). The samples were boiled for 10 min in SDS loading buffer and were then loaded for SDS-polyacrylamide gel electrophoresis (PAGE). After electrophoresis, proteins were transferred from the gel to a polyvinylidene difluoride (PVDF) membrane in a transfer buffer (20 mM Tris, 150 mM glycine). The membrane was rinsed in Tris-buffered saline (TBS) and were then blocked in 5% nonfat dry milk in TBST (TBS with 0.1% Tween 20) for 2 h at 25°C, with 50 rpm shaking, followed by incubation with the mouse anti-HA (Abmart, Shanghai, China) or anti-MYC (Proteintech, Wuhan, China) monoclonal antibody at 4°C overnight. After washing three times with TBST, the membrane was incubated with goat anti-mouse immunoglobulin secondary antibody at 25°C for 1 h. Protein bands were visualized using ECL substrate (Solarbio, Beijing), following manufacturer instructions.

Xu L., Zhang X., Zheng D., Chang Y., Zhang F., Wang Y, & Huang L. (2022). VmMon1-Ccz1 Complex Is Required for Conidiation, Autophagy, and Virulence in Valsa mali. Molecular plant-microbe interactions : MPMI, 35(10).

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