CHLH (GUN5) | Magnesium-chelatase subunit ChlH, chloroplastic

Product no: AS22 4712

AS22 4712   | Clonality:  Polyclonal |  Host:  Rabbit | Reactivity: Hordeum vulgare

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  • Product Info
  • 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
  • Confirmed reactivity: Arabidopsis thaliana, Hordeum vulgare
    Predicted reactivity: Acaryochloris marina, Halomicronema hongdechloris, Synechocystis sp. 6803

    Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Western blot with GUN5 antibodies

    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 extract

    Leaf 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 

  • Background
  • 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
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

    Agrisera Educational Poster Collection
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Buy 2 items of this product for 226.00 €/items
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