Kelch repeat protein (Chlamydomonas)

Name: Kelch repeat protein (Chlamydomonas)
SKU: AS14 2761
Price: 302 EUR
Availability: InStock

AS14 2761 | clonality:polyclonal | host:rabbit | reactivity: Chlamydomonas reinhardtii, Eudorina elegans, Gonium pectorale

Kelch repeat protein (Chlamydomonas) in the group Antibodies Plant/Algal / Plant Developmental Biology / Ion metabolism at Agrisera AB (Antibodies for research) (AS14 2761)

Product Information

Immunogen

Recombinant Cr-Klech, UniProt: A8J2W4, Locus name: g2987 (from Chlamydomonas Reinhardtii)

Host Rabbit

Clonality Polyclonal

Purity Serum

Format Lyophilized

Quantity 50 µl

Reconstitution for reconstitution add 50 µl 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 33,5 kDa

Reactivity

Confirmed reactivity Chlamydomonas reinhardtii (strain 21grm cw15, M10), Gonium pectorale, Eudorina elegans

Predicted reactivity

algae

Species of your interest not listed? Contact us

Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples Application example

Western blot using anti-Kelch antibodies

Total protein from Chlamydomonas reinhardtii (1), Gonium Pectorale (2), Eudorina elegans (3) was extracted using 1x Laemmli buffer + DTT + protease and phosphatase inhibitors without bromophenol blue heated at 95 oC for ten minutes then vortexed with zirconia beads at max speed for ten minutes. Lysate was spun at 4 oC for ten minutes at max speed in a tabletop centrifuge. Proteins were precipitated out of supernatant in 80% acetone at -20 oC for 20 min, spun at 4 oC 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 100 mg chlorophyll/uL. Protein equivalent to 1 ug of chlorophyll was loaded on a 4-20% Bis-Tris gel and blotted to nitrocellulose using wet transfer methods. Blot was blocked with 5% 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/4oC. 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 1:25,000 in 5% milk in TBS-T for 1hr/RT with agitation. The blot was washed as above and developed with chemiluminescent 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

Background

Background Kelch repeat-containing protein (KrP)/Galactose oxidase is a protein associated with iron acquisition and metal homeostasis.

Product citations

immunogen:	Recombinant Cr-Klech, UniProt: A8J2W4, Locus name: g2987 (from Chlamydomonas Reinhardtii)
Reconstitution:	for reconstitution add 50 µl of sterile water
Host:	Rabbit
Clonality:	Polyclonal
Purity:	Serum
Format:	Lyophilized
Quantity:	50 µl
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]:	33,5 kDa
Confirmed reactivity:	Chlamydomonas reinhardtii (strain 21grm cw15, M10), Gonium pectorale, Eudorina elegans
predicted reactivity:	algae Species of your interest not listed? Contact us
not reactive in:	No confirmed exceptions from predicted reactivity are currently known
Picture (footer):	Application example Total protein from Chlamydomonas reinhardtii (1), Gonium Pectorale (2), Eudorina elegans (3) was extracted using 1x Laemmli buffer + DTT + protease and phosphatase inhibitors without bromophenol blue heated at 95 oC for ten minutes then vortexed with zirconia beads at max speed for ten minutes. Lysate was spun at 4 oC for ten minutes at max speed in a tabletop centrifuge. Proteins were precipitated out of supernatant in 80% acetone at -20 oC for 20 min, spun at 4 oC 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 100 mg chlorophyll/uL. Protein equivalent to 1 ug of chlorophyll was loaded on a 4-20% Bis-Tris gel and blotted to nitrocellulose using wet transfer methods. Blot was blocked with 5% 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/4oC. 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 1:25,000 in 5% milk in TBS-T for 1hr/RT with agitation. The blot was washed as above and developed with chemiluminescent 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:	Kelch repeat-containing protein (KrP)/Galactose oxidase is a protein associated with iron acquisition and metal homeostasis.