Anti-TIP1;1, TIP1;2 | tonoplast intrinistic protein 1-1, 1-2 (gamma)

Product no: AS22 4844

AS22 4844 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Oryza sativa

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  • Product Info
  • Immunogen:

    KLH-conjugated synthetic peptide conserved in Raphanus sativus TIP1;1 and TIP1;2 (protein accesion number available in Suga et al. 2001). Peptide is also conserved in Arabidopsis thaliana TIP1-1 P25818, At2g36830, TIP1-2 Q41963, At3g26520

    Host: Rabbit
    Clonality: Polyclonal
    Purity: Antigen affinity purified serum, in PBS pH 7.4
    Format: Lyophilized
    Quantity: 50 µg
    Reconstitution: For reconstitution, add 50 µl of sterile water.
    Storage: Store lyophilized/reconstituted at -20°C; make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
    Tested applications: Western blot (WB)
    Recommended dilution: 1 : 1000 (WB)
    Expected | apparent MW: 25.8 | 23 kDa
  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana, Oryza sativa
    Predicted reactivity: Brassica napus, Gossypium hirsutum, Hordeum vulgare, Populus trichocarpa, Raphanus sativus, Ricinus communis

    Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Western blot using anti-TIP1;1, TIP1§;2 antibodies

    7.5 µg / 37.5 µg and 75 µg of total protein extracted freshly isolated from whole leaf extracts o Arabidopsis thaliana wt plants. Material was grinded frozen and then mixed 1:4 with cold isolation buffer (20 mM Tris pH 7.6, 80 mM NaCl, 0.7 mM EDTA, 5 mM MgCl, 1 mM DTT and 2 % SDS). Protein concentration was measured by Bradford and sample was mixed 1:1 with loading dye (Bromphenolblue with 4 %SDS and 2 % bet-mercaptoethanol). Sample was denaturized at 96 °C for 10 min. Samples were separated on12.5% SDS-PAGE and blotted for 45 min to nitrocellulose, using: semi-dry transfer. Blot was blocked with 5 % milk or for: 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1.000 in TBS-T for ON/4°C with agitation. The antibody solution was decanted, and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, As09 602, Agrisera) diluted to 1: 25.000 in for 1 h at RT with agitation. The blot was washed as above and developed with a following chemiluminescent detection reagent: AgriseraBright (AS16 ECL-N-10). 

    Courtesy of Dr. Andras Bittner, IZMB - Zellbiologie der Pflanzen, University of Bonn, Germany

  • Additional Information
  • Additional information (application):

    Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.

    Diluted antibody solution can be used 2 to 3 times within one month if it contains 0.1 % sodium azide as preservative and is stored at -20ºC to -80ºC.

    Triton X-100 should not be included in the protein extraction buffer, when cell organelles or membrane proteins must be separated from soluble proteins. Because, Triton X breaks membrane structure and solubilizes most membranes proteins. Furthermore, it should be noted that Triton X at high concentrations binds SDS and mask the detergent effect of SDS for SDS-PAGE. Also, micelles of Triton X behave as a large complex with molecular mass of 90 kDa at high concentrations in SDS-PAGE.

  • Background
  • Background:

    TIPs belong to MIP/aquaporin protein family. Alternative names: Aquaporin TIP1-1,gamma-tonoplast intrinsic protein, gamma-TIP, aquaporin TIP, gamma-tonoplast intrinsic protein, gamma-TIP, aquaporin TIP, tonoplast intrinsic protein, root-specific RB7, gamma-tonoplast intrinsic protein 2, gamma-TIP2, salt stress-induced tonoplast intrinsic protein

  • Product Citations
  • Selected references: Mao and Sun (2015). Arabidopsis seed-specific vacuolar aquaporins are involved in maintaining seed longevity under the control of ABSCISIC ACID INSENSITIVE 3. J Exp Bot. 2015 May 26. pii: erv244.
    Suga et al. (2001). Specificity of the accumulation of mRNAs and proteins of the plasma membrane and tonoplast aquaporings in radish organs. Planta 212:294-304.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts


    Oxygenic photosynthesis poster by prof. Govindjee and Dr. Shevela

    Z-scheme of photosynthetic electron transport by prof. Govindjee and Dr. Björn and Dr. Shevela


    Method for isolation of plant vacuolar membranes

     

    protocol for plant vacuolar membrane isolation

    Courtesy of Dr. Masayoshi Maeshima, Laboratory of Cell Dynamics, Graduate School of Bioagricultural Sciences Nagoya University Nagoya, Japan

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