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使用敲除细胞株进行验证重组RabMAb

重组Anti-SAMHD1抗体[EPR13040(B)] - C-terminal (ab177462)

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Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
  • Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
  • Immunocytochemistry/ Immunofluorescence - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
  • Flow Cytometry (Intracellular) - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
  • Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
  • Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR13040(B)] to SAMHD1 - C-terminal
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

概述

  • 产品名称

    Anti-SAMHD1抗体[EPR13040(B)] - C-terminal
    参阅全部 SAMHD1 一抗

  • 描述

    兔单克隆抗体[EPR13040(B)] to SAMHD1 - C-terminal

  • 宿主

    Rabbit

  • 经测试应用

    适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
    不适用于: IP

  • 种属反应性

    与反应: Human

  • 免疫原

    Synthetic peptide within Human SAMHD1 aa 600 to the C-terminus (C terminal) (Cysteine residue). The exact sequence is proprietary.
    Database link: Q9Y3Z3

  • 阳性对照

    • THP1, K562, HeLa and MCF7 cell lysates. Human, spleen and pancreas and tonsil tissue. MCF7 and THP1 cells.

  • 常规说明

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab177462于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
Flow Cyt (Intra)
1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
1/10000 - 1/50000. Predicted molecular weight: 72 kDa.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100 - 1/250.
说明
Flow Cyt (Intra)
1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
1/10000 - 1/50000. Predicted molecular weight: 72 kDa.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100 - 1/250.
应用说明
Is unsuitable for IP.

靶标

  • 功能

    Putative nuclease involved in innate immune response by acting as a negative regulator of the cell-intrinsic antiviral response. May play a role in mediating proinflammatory responses to TNF-alpha signaling.

  • 组织特异性

    Expressed in heart, skeletal muscle, spleen, liver, small intestine, placenta, lung and peripheral blood leukocytes. No expression is seen in brain and thymus.

  • 疾病相关

    Defects in SAMHD1 are the cause of Aicardi-Goutieres syndrome type 5 (AGS5) [MIM:612952]. A form of Aicardi-Goutieres syndrome, a genetically heterogeneous disease characterized by cerebral atrophy, leukoencephalopathy, intracranial calcifications, chronic cerebrospinal fluid (CSF) lymphocytosis, increased CSF alpha-interferon, and negative serologic investigations for common prenatal infection. Clinical features as thrombocytopenia, hepatosplenomegaly and elevated hepatic transaminases along with intermittent fever may erroneously suggest an infective process. Severe neurological dysfunctions manifest in infancy as progressive microcephaly, spasticity, dystonic posturing and profound psychomotor retardation. Death often occurs in early childhood.

  • 序列相似性

    Belongs to the SAMHD1 family.
    Contains 1 HD domain.
    Contains 1 SAM (sterile alpha motif) domain.

  • 细胞定位

    Nucleus.
  • Information by UniProt

  • 数据库链接

    • 别名

      • CHBL2 antibody
      • DCIP antibody
      • Dendritic cell derived IFNG induced protein antibody
      • Dendritic cell-derived IFNG-induced protein antibody
      • Deoxynucleoside triphosphate triphosphohydrolase SAMHD1 antibody
      • dNTPase antibody
      • HD domain containing 1 antibody
      • HDDC1 antibody
      • Mg11 antibody
      • Monocyte protein 5 antibody
      • MOP 5 antibody
      • MOP-5 antibody
      • MOP5 antibody
      • OTTHUMP00000030889 antibody
      • SAM domain and HD domain 1 antibody
      • SAM domain and HD domain containing protein 1 antibody
      • SAM domain and HD domain-containing protein 1 antibody
      • SAMH1_HUMAN antibody
      • Samhd1 antibody
      • SBBI88 antibody
      see all

    图片

    • Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
      Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
      All lanes : Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462) at 1/10000 dilution

      Lane 1 : HEK-293 cell lysate
      Lane 2 : Jurkat cell lysate
      Lane 3 : Wild-type HAP1 cell lysate
      Lane 4 : SAMHD1 knockout HAP1 cell lysate
      Lane 5 : Human Spleen cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 72 kDa
      Observed band size: 73 kDa why is the actual band size different from the predicted?



      Anti-SAMHD1 antibody [EPR13040(B)] (ab177462) staining at 1/10000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab177462 was shown to bind specifically to SAMHD1. A band was observed at 73 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in SAMHD1 knockout cell line. To generate this image, wild-type and SAMHD1 knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    • Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
      Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: SAMHD knockout HAP1 cell lysate (20 µg)
      Lane 3: K562 cell lysate (20 µg)
      Lanes 1 - 3: Merged signal (red and green). Green - ab177462 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab177462 was shown to recognize SAMHD1 when SAMHD1 knockout samples were used, along with additional cross-reactive bands. Wild-type and SAMHD1 knockout samples were subjected to SDS-PAGE. ab177462 and ab8245 (loading control to GAPDH) were both diluted 1/10000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)

      Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling SAMHD1 with ab177462 at 1/100 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
      Immunocytochemistry/ Immunofluorescence - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)

      Immunofluorescence analysis of MCF7 cells labeling SAMHD1 with ab177462 at 1/100 dilution.

    • Flow Cytometry (Intracellular) - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
      Flow Cytometry (Intracellular) - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)

      Intracellular flow cytometric analysis of permeabilized THP1 cells labeling SAMHD1 (red)with ab177462 at 1/10 dilution or rabbit IgG (negative) (green). 

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)

      Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling SAMHD1 with ab177462 at 1/100 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
      Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
      All lanes : Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462) at 1/10000 dilution

      Lane 1 : THP1 cell lysate
      Lane 2 : K562 cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : MCF7 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat-anti-rabbit HRP at 1/2000 dilution

      Developed using the ECL technique.

      Predicted band size: 72 kDa

    • Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)
      Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)

    文献 (0)

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    ab177462 尚未被引用在任何文献中。

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