TKL1 | transketolase (chloroplastic)

AS15 2903 | Clonality: Polyclonal | Host: Rabbit | Reactivity:Arabidopsis thaliana, Pisum sativum

TKL1 | transketolase (chloroplastic) in the group Antibodies for Plant/Algal / Developmental Biology / Ion metabolism at Agrisera AB (Antibodies for research) (AS15 2903)


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product information
Background TKL-1 (Transketolase-1, chloroplastic) is involved in the pathway of Calvin cycle. This enzyme is catalyzing the reversible transfer of a two-carbon ketol group from fructose-6-phopshate or sedoheptuloze-7-phosphate to glyceraldehyde-3-phosphate to yield xylulose-5-phosphate and erythrose-4-phosphate or ribose-5-phosphate. AtTKL1 is expressed in highest levels in photosynthetic tissue, while AtTKL2 is expressed mainly during embryo development. Alternative name: TK.

Purified, recombinant mature TKL1 of Arabidopsis thaliana, UniProt: Q8RWV0, TAIR: At3g60750

Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 ĩl
Reconstitution For reconstitution add 50 ĩl of sterile water.
Storage 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 Immunoprecipitation (IP), Western blot (WB)
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Plant protein extraction buffer

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Additional information

Please notice that this product contains 10% glycerol and might appear as liquid but is provided lyophilized.

application information
Recommended dilution 1 : 2000 (WB)
Expected | apparent MW

72 kDa

Confirmed reactivity Arabidopsis thaliana, Pisum sativum
Predicted reactivity Synechococcus elongatus sp PCC 7942
Not reactive in
Additional information TKL1 has MW of 79.28 kDa with and 73.45 kDa without presequence; about 72 kDa on SDS gel

Antibody works on whole leaf extracts and isolated chloroplasts and is also recognizing recombinant TKL protein.
Selected references Rocha et al. (2014). Phosphorylation of Arabidopsis transketolase at Ser428 provides a potential paradigm for the metabolic control of chloroplast carbon metabolism. Biochem J. 2014 Mar 1;458(2):313-22. doi: 10.1042/BJ20130631.

application example

western blot using anti-TKL antibodies

Approximately 1 ug (in dilution series) of total soluble (stromal) protein of Arabidopsis thaliana extracted from isolated chloroplasts using a buffer containing 10% glycerol, 200 mM NaCl, protease inhibitor and 1mM DTT. Samples were denatured at 96°C for 3 min and were separated on 10 % SDS-PAGE  and blotted for 45min. to PVDF using semi-dry transfer. Blots were blocked with TBST with 5 % milk powder (0.02 % Tween 20)  for 1h at room temperature (RT). Blot was incubated in the primary antibody at a dilution of 1: 5 000 for overnight at 4°C with agitation in TBST with 5 % milk powder. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 10 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 12:0 000 in  for 1h at RT with agitation. The blot was washed as above and developed following manufacture recommendations (SERVALight EOS CL HRP WB Substrate Kit). Exposure time was 10-30 seconds.

Courtesy of Dr. Ute Vothknecht and Edoardo Cutolo, Munich University, Germany


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