Anti-CHLH (GUN5) | Magnesium-chelatase subunit ChlH, chloroplastic
AS22 4712 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Hordeum vulgare
- Product Info
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Immunogen: KLH-conjugated peptide derived from Arabidopsis thaliana CHLH protein sequence, UniProt: Q9FNB0, TAIR: At5g13630 Host: Rabbit Clonality: Polyclonal Purity: Antigen affinity purified in PBS pH 7.4 Format: Lyophilized Quantity: 50 µg Reconstitution: For reconstitution, add 50 µl of sterile or deionized 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: Western blot (WB) Recommended dilution: 1 : 5000 (WB) Expected | apparent MW: 153.57 | 130 kDa - Reactivity
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Confirmed reactivity: Arabidopsis thaliana, Hordeum vulgare Predicted reactivity: Acaryochloris marina, Halomicronema hongdechloris, Synechocystis sp. 6803
Species of your interest not listed? Contact usNot reactive in: No confirmed exceptions from predicted reactivity are currently known - Application Examples
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Samples:
1 - 50 µg Hordeum vulgare whole leaf extract
2 - 50 µg Hordeum vulgare gun5 knock out whole leaf extract
3 - 30 µg Hordeum vulgare whole leaf extract
4 - 30 µg Hordeum vulgare gun5 knock out whole leaf extract
5 - 10 µg Hordeum vulgare whole leaf extract
6 - 10 µg Hordeum vulgare gun5 knock out whole leaf extractLeaf material was homogenized in Laemmli sample buffer [20% (v/v) glycerol, 4% (w/v) SDS, 160 mM Tris–HCl pH 6.8, 10% (v/v) 2-mercaptoethanol] to a concentration of 0.1 mg µl−1 (fresh weight/Laemmli sample buffer). Samples were incubated at 65°C for 15 min and, after a centrifugation step (10 min at 16 000 g), the supernatant was incubated for 5 min at 95°C and loaded onto 10 % SDS-PAGE Acrylamide/Bis-acrylamide (29:1) and blotted for 1 h to PVDF (0.45 µm pore size), using semi-dry or dry transfer (Biorad). Blot was blocked with 5 % milk for: 4h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5000 for ON/4°C with agitation. The antibody solution was decanted, and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated AS09 602, Agrisera) diluted to 1: 25 000 in for h/RT with agitation. The blot was washed as above and developed with a following chemiluminescent detection reagent: AS16 ECL-N-10 AgriseraBright (mid picogram). Exposure time was 2 minutes.
Courtesy of Dr. Luca Tadini, Università degli studi di Milano, Italy
Samples:
1 - 10 ug of 4 days old of wild-type Arabidopsis thaliana seedlings extract
2 - 5 ug of 4 days old of wild-type Arabidopsis thaliana seedlings extract
3- 10 ug of 4 days old of gun5-1 mutant seedlings extract
Mark: MW markers
Target MW: 153.57 kDa
5-10 µg/well of total protein extracted from fresh 4 days old of Arabidopsis thaliana whole seedling. Exact buffer components were: (50 mM Tris–HCl pH 7.5, 10% glycerol, 150 mM NaCl, 10 mM MgCl2, 5 mM EDTA, 5 mM DTT, 0.5% (v/v) Triton X-100, and 1 × protease inhibitors) and denatured with 4X SDS sample loading buffer (200 mM Tris-Cl (pH 6.8). 8% SDS (sodium dodecyl sulfate). 0.4% Bromophenol blue. 40% glycerol) at 95°C 10 min. Samples were separated in the RT on 10 % SDS-PAGE and blotted for 0.5 h to PVDF (pore size of 0.45 um), using: semi-dry at room temperature. Blot was blocked with 5 % milk for: 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2 000 at 4°C with agitation overnight. The antibody solution was decanted, and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602 Agrisera) diluted to 1: 50 000 for 2 h/RT with agitation. The blot was washed as above and developed with a following chemiluminescent detection reagent: AS16 ECL-N-10 AgriseraBright (mid picogram). Exposure time was 2 minutes.
Courtesy of Dr. Duorong Xu, LMU München, Germany
- Background
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Background: CHLH (GUN5) (Magnesium-chelatase subunit ChlH, chloroplastic) is a protein involved in chlorophyll synthesis, plastid to nucleus retrograde signaling and ABA perception.
Alternative names: ABA-binding protein, Mg-protoporphyrin IX chelatase subunit ChlH, Protein CONDITIONAL CHLORINA, Protein GENOMES UNCOUPLED 5, Protein RAPID TRANSPIRATION IN DETACHED LEAVES 1 - Protocols
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Agrisera Western Blot protocol and video tutorials
Protocols to work with plant and algal protein extracts
Agrisera Educational Poster Collection - Reviews:
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Accessories
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Benefits of using this antibody
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Benefits of using this antibody