2 | Rubisco quantitation kit

533 €

AS09 409 | clonality:polyclonal | host: rabbit | r eactivity: dicots, monocots, algae, cyanobacteria


81 st
Item No:
AS09 409

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product information

The Rubisco quantitation kit is suitable for quantitation of Rubisco in plant and algal samples using quantitative immunoblotting. It contains anti-RbcL antibody, Rubisco protein standard and the Protein Extraction Buffer for quantitative protein extraction. This kit is a good alternative to quantification of Rubisco by ELISA as it allows separation of Rubisco from a total complex cell protein mixture, which can be troublesome in case of ELISA method.

Immunogen KLH-conjugated synthetic peptide was used to elicit anti-RbcL antibody. No baculovirus was used in production of this product.
Host Rabbit. Secondary antibody: Goat.
Clonality Polyclonal
Quantity 1 x 50 ĩl of AS03 037, RbcL | Rubisco large subunit, form I and form II (amount enough for 50-100 Western blots)1 x 100 ĩl of AS01 017S, Rubisco protein standard (0.15 pmoles / ĩl, amount enough for generation of standard curve in 85assays (standard curve: 0.0625 pmoles, 0.125 pmoles, 0.25 pmoles)2 x 2 ml AS08 300, Protein extraction buffer (amount enough for 48 isolations of plant material, using 500 ĩl 1x PEB for 100 mg fresh weight) or 120 isolations of algal material (using 200 ĩl 1x PEB for cell amounts corresponding to 4-10 ĩg total chlorophyll)1 x 50 ĩl of AS09 602, Goat anti-rabbit IgG (H&L), HRP conjugated(amount enough for 50-100 Western blots)

RbcL antibody and protein standard:
Please stoore at -20°C (6 month) or -80°C 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 +4°C and for long term storage (1 year) at -20°C.

Tested applications Western blot (WB)
Related products

AS03 037 | anti-RbcL | Rubisco large subunit, form I and form II (50 µl)
AS03 037A | anti-RbcL | Rubisco large subunit, form I and form II (50 µg affinity purified)

AS03 037-HRP| anti-RbcL | Rubisco large subunit, form I and form II (40 µg, HRP-conjugated)

AS15 2994 | Rubisco ELISA quantitation kit 

AS15 2955
| anti-RbcL II | Rubisco large subunit, form II (50 µl), rabbit antibody
AS15 2955S | RbcL II | Rubisco form II positive control/quantitation standard

AS01 017  | anti-RbcL | Rubisco large subunit, form I, chicken antibody
AS01 017S | Rubisco protein standard for quantitative western blot or positive control

AS03 037PRE | Rubisco large subunit, pre-immune serum

AS07 218  | anti-Rubisco | 557 kDa hexadecamer, rabbit antibody to a whole protein

AS07 259
| anti-RbcS | Rubisco small subunit (SSU), rabbit antibody
AS07 222
| anti-RbcS | Rubisco small subunit (SSU) from pea, rabbit antibody

matching secondary antibody

Additional information Recommended secondary antibody: AS09 602, Goat anti-rabbit IgG (H&L), HRP conjugated
application information
Recommended dilution

1:5000-1:10000 with standard ECL (WB)

Expected | apparent MW

Rubisco protein standard: 52.7 kDa (Arabidopsis thaliana), 52.5 kDa (cyanobacteria), 52.3 (Chlamydomonas reinhardtii)

Confirmed reactivity

Arabidopsis thaliana, Chlamydomonas reinhardtii, Cyanophora paradoxa, Emiliana huxleyi, Euglena gracilis, Gonyaulax polyedra, Heterosigma akashiwo, Micromonas pusila, Porphyra sp. Spinacia oleracea, Synechococcus PCC 7942, Thalassiosira pseudonana

Predicted reactivity

di and monocots, conifers, mosses, liverworts, welwitschia, green algae, red alge, brown algae, cryptomonad, cyanobacteria including prochlorophytes, gamma-proeobacteria, beta-proteobacteria, alpha proteobacteria

Not reactive in
Additional information

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.

Selected references Mota et al. (2015). Effects of heavy metals on Cyanothece sp. CCY 0110 growth, extracellular polymeric substances (EPS) production, ultrastructure and protein profiles. J Proteomics. 2015 Apr 29;120:75-94. doi: 10.1016/j.jprot.2015.03.004.
Thamatrakoln et al. (2013). Death-specific protein in a marine diatom regulates photosynthetic responses to iron and light availability. Proc Natl Acad Sci U S A. 2013 Dec 10;110(50):20123-8. doi: 10.1073/pnas.1304727110. Supplemental material describes western blot quantification method.

application example

2 µg of total protein from various plant extracts (1-5) extracted with 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.

For more detailed information check here.


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