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Pan RAB (Internal Region) 抗体

ABIN572061 产品详细信息, 供应商: Log in to see
抗原
抗原表位
Internal Region
适用
人, 小鼠, 大鼠
宿主

克隆类型
多克隆
应用范围
Immunohistochemistry (IHC), Western Blotting (WB)
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Available images

'独立验证'标志
抗原 RAB
Lot Number Rb1082-020809-WS
Method validated Immunocytochemistry
Positive Control HeLa
Negative Control no primary or no secondary antibody
Notes ABIN572061 was successfully validated for ICC based the staining pattern consistent with published data.
Primary Antibody ABIN572061
Secondary Antibody goat anti-rabbit IgG (H+L) secondary antibody, Alexa Fluor 546 (ThermoFisher, A-11035)
实验流程
  • Grow HeLa cells in DMEM growth medium at 37°C in 5% CO2 to 70% confluency.
  • Fix cells in warm 4% formaldehyde for 20min at RT.
  • Wash cells twice with cold PBS.
  • Block and permeabilize cells in B-PBST (PBS, 1% BSA, 0.25% Triton X-100, 0.22µm filter sterilized) for 30min at RT.
  • Incubation with primary antibody ABIN572061 (antibodies-online, lot Rb1082-020809-WS) diluted 1:50 and 1:200 in B-PBST for 1h at RT.
  • Wash cells twice with PBS for 2min at RT.
  • Incubation with goat anti-rabbit IgG (H+L) secondary antibody, Alexa Fluor 546 (ThermoFisher, A-11035) diluted 1:500 in B-PBS for 1h art RT in the dark.
  • Wash cells thrice for 2min at RT in the dark.
  • Mount in VECTASHIELD HardSet Antifade Mounting Medium with DAPI (Vector laboratories, H-1500, lot ZC0816).
  • Confocal imaging on Zeiss LSM 880 at 63x magnification.
    Experimental Notes
    • ABIN572061 works very well in HeLa cells at both dilutions tested, 1:50 & 1:200. The strength of the signal at the 1:200 dilution suggests that it can be diluted even higher to 1:500 or 1:1000 without major loss in image quality.
    • A rabbit anti-giantin antibody in combination with the same secondary antibody as described in the protocol showed the expected localization. Also, both negative controls – no primary or no secondary antibody control – did not show any staining, indicating that the staining pattern shown in the image was indeed due to specific binding of ABIN572061 to its antigen(s).
    Validation Images
    Immunocytochemistry validation image for anti-Pan RAB (Internal Region) antibody (ABIN572061) Staining of HeLa cells with ABIN572061 in conjunction with an AF546 conjugated goat a...
    免疫原 A synthetic peptide from internal region of mouse Ras-related protein Rab-3A (RAB3A) conjugated to an immunogenic carrier protein has been used as the antigen. The antigen is homologous or shares a significant identity with many RAB molecules including RAB1, RAB2, RAB4, RAB9, RAB10, RAB11, RAB14, RAB26, RAB30, RAB35, RAB39, RAB43.
    特异性 Specific for RAB.
    纯化方法 Whole serum
    背景 Function: Involved in exocytosis by regulating a late step in synaptic vesicle fusion. Could play a role in neurotransmitter release by regulating membrane flow in the nerve terminal.
    Subcellular location: Cell membrane, Lipid-anchor, Cytoplasmic side Also known as: RAB3, RAB4, RAB9, RAB43, RAB35, RAB39, RAB30, RAB1, RAB4, RAB14, RAB11, RAB10, RAB26, RAB2.
    应用备注 Use at a dilution of 1:300 to 1: 2000.
    The optimal dilution should be determined by the end user.
    Not yet tested in other applications.
    限制 仅限研究用
    状态 Lyophilized
    溶解方式 Reconstitute in 100 µL of sterile water. Centrifuge to remove any insoluble material.
    注意事项 Avoid freeze and thaw cycles.
    储存条件 4 °C/-20 °C
    储存方法 Maintain the lyophilised/reconstituted antibodies frozen at -20°C for long term storage and refrigerated at 2-8°C for a shorter term. When reconstituting, glycerol (1:1) may be added for an additional stability. Avoid freeze and thaw cycles.
    有效期 12 months