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Anti-HSP60/HSPD1 Antibody (Clone#6G2)

Mouse monoclonal antibody

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筛选器: All WB IHC FCM ICC/IF

M01280-3

  • 50μl ¥1180 100μl ¥1960 150μl ¥2780
  • 货期: 现货
  • Figure 1. Western blot analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
    Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.

    Lane 1: human Caco-2 whole cell lysates

    Lane 2: human A549 whole cell lysates

    Lane 3: human THP-1 whole cell lysates

    Lane 4: human SW620 whole cell lysates

    Lane 5: human U-937 whole cell lysates

    Lane 6: human HepG2 whole cell lysates

    Lane 7: rat RH35 whole cell lysates

    Lane 8: mouse RAW246.7 whole cell lysates
    After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-HSPD1 antigen affinity purified monoclonal antibody (Catalog # M01280-3) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for HSPD1 at approximately 60KD. The expected band size for HSPD1 is at 60KD.

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  • Figure 2. IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
    HSPD1 was detected in paraffin-embedded section of human intestinal cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-HSPD1 Antibody (M01280-3) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.

    all(9)
  • Figure 3. IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
    HSPD1 was detected in paraffin-embedded section of human lung cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-HSPD1 Antibody (M01280-3) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.

    all(9)
  • Figure 4. IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
    HSPD1 was detected in paraffin-embedded section of human mammary cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-HSPD1 Antibody (M01280-3) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.

    all(9)
  • Figure 5. IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
    HSPD1 was detected in paraffin-embedded section of mouse liver tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-HSPD1 Antibody (M01280-3) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.

    all(9)
  • Figure 6. IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
    HSPD1 was detected in paraffin-embedded section of rat liver tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-HSPD1 Antibody (M01280-3) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.

    all(9)
  • Figure 7. Flow Cytometry analysis of A431 cells using anti-HSPD1 antibody (M01280-3).
    Overlay histogram showing A431 cells stained with M01280-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-HSPD1 Antibody (M01280-3, 1:100) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 1:100) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1:100) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

    all(9)
  • Figure 8. Flow Cytometry analysis of HepG2 cells using anti-HSPD1 antibody (M01280-3).
    Overlay histogram showing HepG2 cells stained with M01280-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-HSPD1 Antibody (M01280-3, 1:100) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 1:100) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1:100) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

    all(9)
  • Figure 9. ICC analysis using anti- HSPD1 antibody (M01280-3). was detected in immersion fixed A431 cell line. Cells were stained using the Dylight488-conjugated Anti-mouse IgG Secondary Antibody (green)(Catalog # BA1126) and counterstained with DAPI (blue).

    all(9)

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产品名称
Anti-HSP60/HSPD1 Antibody (Clone#6G2)
规格/价格
50μl/1180 100μl/1960 150μl/2780
指标别称
HSP 60; HSP60; HSP65; HSPD1
产品类型
Monoclonal
检验物种
human, mouse, rat
应用范围
WB, IHC, ICC/IF, FCM
基因名称
HSPD1
克隆号
6G2
宿主
Mouse
抗体亚型
IgG1
免疫原
E.coli-derived human Hsp60/HSPD1 recombinant protein (Position: A260-Q496). Human Hsp60 shares 97% amino acid (aa) sequence identity with both mouse and rat Hsp60.
计算分子量
61 kDa
实际分子量
60 kDa
成分
500 ug/ml antibody with PBS, 0.02% NaN3, 1 mg/ml BSA and 50% glycerol.
纯化方式
protein G purified.
浓度
500 ug/ml
产品形态
Liquid
保存条件
12 months from date of receipt,-20℃ as supplied. 6 months 2 to 8℃ after reconstitution. Avoid repeated freezing and thawing.
背景资料
HSP60 is a member of the chaperonin class of protein factors, which include the Escherichia coli groEL protein and the Rubisco subunit-binding protein of chloroplasts. It acts as a costimulator of human regulatory CD4-positive/CD25 -positive T cells, which inhibit lymphoproliferation and IFNG and TNF secretion by CD4-positive and CD8-positive T cells. HSP60 enhances Treg activity via TLR2, leading to activation of an intracellular signaling cascade that included p38, as well as inhibition of ERK phosphorylation. Suppression of target T cells is mediated by both cell-to-cell contact and by secretion of TGFB and IL10, and it leads to downregulation of ERK, NFKB, and TBET expression. The self-molecule HSP60 can downregulate adaptive immune responses by upregulating Tregs through TLR2 signaling.
Uniprot ID
P10809  
RRID
文献引用格式
HSP60/HSPD1 Antibody (Clone#6G2) (Boster Biological Technology, Wuhan, China. Catalog#M01280-3)
应用释义
WB- 蛋白质免疫印迹法,IHC- 免疫组织化学法,ICC/IF-免疫细胞荧光,ICC-免疫细胞化学,IHC-F- 冰冻切片免疫组化,FCM-流式细胞术,ELISA-酶联免疫吸附测定,IP-免疫沉淀法 ,IF-免疫组织荧光法,ChIP-染色质免疫沉淀法
推荐配套的二抗和检测试剂
Boster recommends Enhanced Chemiluminescent Kit with anti-Mouse IgG (EK1001) for Western blot, and HRP Conjugated anti-Mouse IgG Super Vision Assay Kit (SV0001-1) for IHC(P) and ICC.
推荐稀释比
Western blot (WB):1:500-2000
Immunohistochemistry (IHC): 1:50-400
Immunocytochemistry/Immunofluorescence (ICC/IF):1:50-400
Flow Cytometry (Fixed):1:50-200
(Boiling the paraffin sections in 10mM citrate buffer,pH6.0,or PH8.0 EDTA repair liquid for 20 mins is required for the staining of formalin/paraffin sections.) Optimal working dilutions must be determined by end user.

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    Anti-HSP60/HSPD1 Antibody (Clone#6G2)

    筛选器: All WB IHC FCM ICC/IF

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