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HXK1 | Hexokinase 1 (Chlamydomonas)

AS16 4083  |  Clonality: Polyclonal   |  Host: Rabbit  |  Reactivity: Chlamydomonas reinhardtii, Eudorina elegans, Gonium pectorale

HXK1 | Hexokinase 1 (Chlamydomonas) in the group Antibodies Plant/Algal  / Plant Developmental Biology / Plant Signal Transduction at Agrisera AB (Antibodies for research) (AS16 4083)
HXK1 | Hexokinase 1 (Chlamydomonas)



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How to cite this product:
Product name, number (Agrisera, Sweden)

Data sheet Product citations Protocols Add review

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Chlamydomonas reinhardtii hexokinase-1, UniProt: A8JGU7

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 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 67,3 kDa

Reactivity

Confirmed reactivity Chlamydomonas reinhardtii, Eudorina elegans, Gonium pectorale
Predicted reactivity
Species of your interest not listed? Contact us

Application examples

Application examples Application example

Western blot using anti-HXK1 (Chlamydomonas)


Total protein of Chlamydomonas reinhardtii strain 21gr (1), strain cw15 (2), strain M10 (3) was extracted using 1x Laemmli buffer + DTT + protease and phosphatase inhibitors without bromophenol blue heated at 95°C for ten minutes then vortexed with zirconia beads at max speed for ten minutes. Lysate was spun at 4°C for ten minutes at max speed in a tabletop centrifuge. Proteins were precipitated out of supernatant in 80% acetone at -20°C for 20 min, spun at 4°C at max speed for 10 min. Pellet was washed 1x with 80% acetone and resuspended in 1x LDS sample buffer + DTT + inhibitors at a concentration of 1 µg chlorophyll/uL. Protein equivalent to1 of chlorophyll was loaded on a 4-20% Bis-Tris gel and blotted to PVDF using wet transfer methods. Blot was blocked with 10% milk in TBS-T for 1hr/RT. Blot was incubated in the primary antibody at a dilution of 1:1000 in 5% milk in TBS-T ON/4°C. The antibody solution was decanted and the blot was rinsed 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary diluted to 1:50 000 (AS09 602)  in 5% milk in TBS-T for 1hr/RT with agitation. The blot was washed as above and developed withchemiluminescent detection reagent and imaged on a Licor C-Digit blot scanner. Exposure time was 12 min.

Courtesy of Jessica Rakijas, Kansas State University, USA

Additional information

Related products

Background

Background

HXK1 (Hexokinase 1) is an enzyme of plant glucose-signaling network which functions as a glucose sensor. Alternative name: Protein GLUCOSE INSENSITIVE 2.

Product citations

Selected references Upadhyaya and Jagadeeshwar Rao (2019). Reciprocal regulation of photosynthesis and mitochondrial respiration by TOR kinase in Chlamydomonas reinhardtii. Plant Direct Volume 3, Issue 11.
immunogen:

KLH-conjugated synthetic peptide derived from Chlamydomonas reinhardtii hexokinase-1, UniProt: A8JGU7

Reconstitution: For reconstitution add 50 ĩl of sterile water
Host: Rabbit
Clonality: Polyclonal
Purity: Serum
Format: Lyophilized
Quantity: 50 ĩl
storage: Store lyophilized/reconstituted at -20°C; once reconstituted 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)

calculated | apparent molecular mass [kDa]: 67,3 kDa
Confirmed reactivity: Chlamydomonas reinhardtii, Eudorina elegans, Gonium pectorale
predicted reactivity:
Species of your interest not listed? Contact us
Picture (footer): Application example

Western blot using anti-HXK1 (Chlamydomonas)


Total protein of Chlamydomonas reinhardtii strain 21gr (1), strain cw15 (2), strain M10 (3) was extracted using 1x Laemmli buffer + DTT + protease and phosphatase inhibitors without bromophenol blue heated at 95°C for ten minutes then vortexed with zirconia beads at max speed for ten minutes. Lysate was spun at 4°C for ten minutes at max speed in a tabletop centrifuge. Proteins were precipitated out of supernatant in 80% acetone at -20°C for 20 min, spun at 4°C at max speed for 10 min. Pellet was washed 1x with 80% acetone and resuspended in 1x LDS sample buffer + DTT + inhibitors at a concentration of 1 µg chlorophyll/uL. Protein equivalent to1 of chlorophyll was loaded on a 4-20% Bis-Tris gel and blotted to PVDF using wet transfer methods. Blot was blocked with 10% milk in TBS-T for 1hr/RT. Blot was incubated in the primary antibody at a dilution of 1:1000 in 5% milk in TBS-T ON/4°C. The antibody solution was decanted and the blot was rinsed 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary diluted to 1:50 000 (AS09 602)  in 5% milk in TBS-T for 1hr/RT with agitation. The blot was washed as above and developed withchemiluminescent detection reagent and imaged on a Licor C-Digit blot scanner. Exposure time was 12 min.

Courtesy of Jessica Rakijas, Kansas State University, USA
background:

HXK1 (Hexokinase 1) is an enzyme of plant glucose-signaling network which functions as a glucose sensor. Alternative name: Protein GLUCOSE INSENSITIVE 2.

All references: Upadhyaya and Jagadeeshwar Rao (2019). Reciprocal regulation of photosynthesis and mitochondrial respiration by TOR kinase in Chlamydomonas reinhardtii. Plant Direct Volume 3, Issue 11.

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