2 | Educational antibody kit - photosynthesis

1236 €

AS05 070 | clonality: polyclonal | host: rabbit | reactivity: dicots, monocots, algae


93 st
Item No:
AS05 070

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product information

 This kit contains antibodies to the following 5 proteins and 4 protein standards for their quantification. It also includes a matching secondary antibody.

- universal and absolutely essential carbon fixation enzyme that is conserved across oxygenic photoautotrophs and other groups. The RbcL (Rubisco large subunit) protein is hyperabundant and regulation of RbcL content under different conditions or stress responses sets a limit on the maximum capacity for CO2 uptake in a population or community.

Proteins involved in conversion of solar energy into chemical bonds:

protein of multisubunit protein complex - Photosystem I, which harnesses light energy required for photosynthesis to occur.

PsbA protein of multisubunit complex - Photosystem II -  the ultimate source of almost all biosynthetic reductant in the biosphere. PsbA (D1) protein is rapidly cycled under illumination in all oxygenic photobionts. Tracking PsbA pools using the Global PsbA Antibody shows the functional content of Photosystem II in a wide range of samples.

Lhcb1 - harnesses light energy for photosynthesis (Photosystem II)

AtpB highly conserved across the beta subunits of known F-type ATP Synthases from chloroplasts, mitochondria and most bacteria. The ATP Synthase complex is essential for the synthesis of ATP from ADP and free phosphate


KLH-conjugated respective synthetic peptides were used

Host Rabbit. Secondary antibody: Goat.
Clonality Polyclonal
Quantity 10 or 50 ĩl / tube of each antibody. 100 or 250 ĩl / tube of each protein standard. 50 ĩl of secondary antibody

store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Tested applications
Related products

Plant and algal protein extraction buffer

Secondary antibodies

Additional information This kit can be used during teaching labs and contains following antibodies and protein standards:

Product information - Primary antibodies:

Product number:

Product name:


Recommended dilution:

AS01 004

Anti-Lhcb1 antibody*

Please follow the instructions on the lable on each vial.

1:2000 for ECL detection.

AS03 037

Anti-RbcL global antibody*

Please follow the instructions on the lable on each vial.

1:5000-10 000 for ECL detection.

AS10 939

Anti-PsaC global antibody*

Please follow the instructions on the lable on each vial.

1:1000 for ECL detection.

AS05 084

Anti-PsbA antibody*

Please follow the instructions on the lable on each vial.

1:10 000 for ECL detection.

AS05 085-10

Anti-AtpB antibody*

Please follow the instructions on the lable on each vial.

1:2000-5000 for ECL detection.

* All primary antibodies in this kit are raised in rabbits.

Product information - Protein standards/positive controls:

Product number:

Product name:


Amount to load on the gel:


AS01 016S

PsbA protein standard

Please follow the instructions on the lable on each vial.

2 μl (0.25 pmol/ μl)

41.5 kDa*

AS01 017S

RbcL (Rubisco) protein standard

Please follow the instructions on the lable on each vial.

2 μl (0.056 pmol/ μl)

52.7 kDa

AS03 030S

AtpB protein standard

Please follow the instructions on the lable on each vial.

10 μl (0.27 pmol/μl)

53.1 kDa*

AS04 042S

PsaC protein standard

Please follow the instructions on the lable on each vial.

2 μl (0.15 pmol/μl)

11.5 kDa*

*These proteins are larger than a respective native protein due to the addition of His-tag

Product information - Secondary antibodiy:

AS09 602- Goat anti-Rabbit IgG (H&L), HRP conjugated, 50 µl (2x25 µl)


Educational information about Quantitative western blot can be found here:  detailed method description, video tutorial

application information
Recommended dilution
Expected | apparent MW
Confirmed reactivity
Predicted reactivity
Not reactive in
Additional information

Suggested experimental treatments for student labs:

- Low light vs. High light
Shift from low or moderate light (50-100 umol photons m-2 s-1) to high light (500-2000 umol photons m-2 s-1, or nearly full sunlight).
Note: Depending on the species and the developmental stage this will lead to large changes in protein profiles, usually within a few hours and is reflected in detectable differences in LHC & RbcL content, and also differences in the PsbA/PsaC ratio (PSII:PSI).
-PsbA breakdown products will become more prominent, and the pool of mature PsbA usually declines
-there may be declines in antenna proteins, and sometimes increases in RbcL, if the stress is not too harsh.
For higher crop plants, it is probably the most generally reliable, rapid way to provoke detectable changes in protein content, since the PsbA/D1 pool will usually decline.

- Changes in nutritional status
Shifting from nutrient replete to low-nutrient conditions will cause gradual declines in LHC & RbcL content.
Etiolation vs. greening
Characteristic change, usually PSI (PsaC) accumulates before PSII (PsbA) and LHCII can be observed.
Developmental stages
Newly emerged, young & old leaves will show different profiles, as will (of course) leaves & roots.

Suggested material:
Plants (spinach, sunflower, pea), Algae or cyanobacteria.

Material preparation and extraction:
Tissue samples of the size required for later extraction can be frozen in foil sachets and stored at –80ºC.
Extraction buffer:
LiDS is recommended over SDS for extraction involving cold steps.
Protease inhibitor: PEFABLOC. For a teaching lab, this may not be strictly necessary, however
if extracts are to be refrozen and reused, degradation becomes an issue and addition of protease inhibitor is advised.
Recommended extraction volume: 150 mg of leaf tissue into 500 ul of extraction buffer (see appendix). After the extraction by grinding and/or sonication the mixture is centrifuged (full speed in microfuge for 2 min) to separate solubilized extract from bulk leaf material. The recovery of extract should be about 300 ul, depending on the type of leaves. This gives an extract at a concentration of roughly 100 ug chl/ml (0.1 ug chl/ul).
Load per well: ca. 0.5 ug chl (so ca. 5 ul of the extract).
So 150 mg of leaf tissue should give a recovered extract of 300 ul @ 0.1 ug chl/ul = 30 ug chl, sufficient for about 60 lane loads of 0.5 ug chl/lane for immunoblotting
The loads can be reduced easily down to 0.2 ug chl (for mini gels) or even lower (down to 0.01 ug chl in some cases using ultrasensitive detection methods). Lighter loads are recommended for some abundant proteins detected with strong antibodies (like anti-RbcL).
Alternatively, for ease of extraction it can be scaled up to 1 g of tissue in 4 ml of extraction buffer. Loads can be prepared based on: chlorophyll, protein or leaf area.
Chlorophyll is suitable and simple loading parameter for determination of the level of the photosynthetic apparatus, which accounts for most leaf protein. Therefore loading by equal chlorophyll can show both qualitative and quantitative changes in a protein profile. For leaves, loads by equal area are recommended, as a leaf is nearly 2-dimensional and photosynthetic capacity per leaf area is a useful ecophysiological parameter.

Spectrophotometric chl determination
Extract chl from ca. 100 ul of aqueous extract into 900 ul of 80% acetone saturated with MgCO3.
Measure at A663 and A750 (Blank=1 ml 80 % acetone).
[Chla](μ = 12.7 (A663-A750)x 1000/x

It is recommended to run 3 lanes of quantitated standard to generate a standard curve. It can be helpful to perform trial runs to get the quantitation standards and samples in the same range. Immunoblottting with primary and secondary antibodies and ECL shows linearity of detection over about 1 order of magnitude. Bands are detected over a much wider range, but the pseudo-linearity load/response region is only about 10 fold.

Agrisera Western blot protocols and additional information about antibodies

Selected references

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