IKP

Rubisco quantitation kit (Western blot)

Product no: AS09 409set
AS09 409set | Clonality: Polyclonal | Host: Rabbit

This set enables quantification of Rubisco using Western blot in samples from a range of various species  including dicots, monocots, algae, cyanobacteria. Benefits of using this antibody set
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Delivery 3-6 business days
Delivery:  3-6 business days
  • Data sheet

  • This set contains the following products:

    AS03 037, RbcL | Rubisco large subunit, form I and form II (50 µl)
    AS01 017S, Rubisco protein standard (100 µl)
    AS08 300, Protein extraction buffer (2 x 2 ml)
    AS09 602, Goat anti-rabbit IgG (H&L), HRP conjugated(2 x 10 µl)
    AS16 ECL-N-10, AgriseraECL Bright (10 ml)

    • Product Info
    • Immunogen: KLH-conjugated synthetic peptide was used to elicit anti-RbcL antibody. No baculovirus was used in production of this product.
      Host: Primary antibody: Rabbit, Secondary antibody: Goat,
      Clonality: Polyclonal
      Quantity: 1 x 50 l of AS03 037, RbcL | Rubisco large subunit, form I and form II (amount enough for 50-100 Western blots)1 x 100 l of AS01 017S, Rubisco protein standard (0.15 pmoles / l, amount enough for generation of standard curve in 34 assays (standard curve: 0.0625 pmoles, 0.125 pmoles, 0.25 pmoles)2 x 2 ml of AS08 300, Protein extraction buffer (amount enough for 48 isolations of plant material, using 500 l 1x PEB for 100 mg fresh weight) or 120 isolations of algal material (using 200 l 1x PEB for cell amounts corresponding to 4-10 g total chlorophyll)2 x 10 l of AS09 602, Goat anti-rabbit IgG (H&L), HRP conjugated(amount enough for 50-100 Western blots)1 X 10 ml of AS16 ECL-N-10, AgriseraECL Bright 10 ml (5 ml of component A + 5 ml of component B, trial pack)
      Storage: RbcL antibody and protein standard: Please store at -20C (6 months) or -80C for long term storage(years). Please avoid freezing and thawing of reconstituted antibodies, make aliquots instead. PEB extraction buffer:Stable at RT for at least 1 month. For short-term storage please stoore (1 month) at 4C and for long term storage (1 year) at -20C.
      Tested applications: Western blot (WB)
      Recommended dilution:
      1 : 5000-1 : 10000 (WB)
      Expected | apparent MW: Rubisco protein standard: 52.7 kDa (Arabidopsis thaliana), 52.5 kDa (cyanobacteria), 52.3 (Chlamydomonas reinhardtii)
    • Reactivity
    • Confirmed reactivity: Arabidopsis thaliana, Chlamydomonas reinhardtii, Cyanophora paradoxa, Cynara cardunculus var altilis, Emiliana huxleyi, Euglena gracilis, Gonyaulax polyedra, Heterosigma akashiwo, Micromonas pusila, Porphyra sp. Spinacia oleracea, Synechococcus PCC 7942, Thalassiosira pseudonana
      Predicted reactivity: Alpha proteobacteria, Algae (brown and red), Beta-proteobacteria, Dicots, Conifers, Cryptomonad, Cyanobacteria, Gamma-proeobacteria, Liverworts, Mosses, Prochlorophytes, Pelwitschia
      Species of your interest not listed? Contact us
    • Application Examples
    • Application example


      example

      2 µg of total protein from various plant extracts (1-5) extracted with Agriera Protein Extraction Buffer PEB (AS08 300) separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Markers MagicMarks (Invitrogen) (M) and Rubisco protein standard (AS01 017S) at 0.0625 pmol, 0.125 pmol, 0.25 pmol.

      Following standard western blot procedure this image has been obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad). The contour tool of the software is used to the area for quantitation and the values are background subtracted to give an adjusted volume in counts for each standard and sample.

      Agrisera Western Blot protocol and video tutorials

    • Additional Information
    • Additional information (application): Quantitative western blot:  detailed method description, video tutorial

      Discussion over some critical aspects of quantitative western blot: Heidebrecht et al. (2009). Improved semiquantitative Western blot technique with increased quantification range. J. Immunological Methods. 35:40-48.
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