Hi Preeti,
I would say that you have different oligomers in your resin after the whole
process. 72 kDa would be a heterodimer (L+H), whereas 95 would be a homodimer
(H+H).
Best wishes
______________________________________________________
Rafael Marques da Silva
PhD Student – Structural Biology
University of Leicester
Mestre em Física Biomolecular
Universidade de São Paulo
Bacharel em Ciências Biológicas
Universidade Federal de São Carlos
phone: +44 07861 273773
"A sorte acompanha uma mente bem treinada"
________________________________________________
________________________________
De: CCP4 bulletin board <[email protected]> em nome de Preeti
<[email protected]>
Enviado: quarta-feira, 13 de agosto de 2025 17:43
Para: [email protected] <[email protected]>
Assunto: [ccp4bb] Unusual SDS-PAGE pattern from IgG beads
Hi everyone,
I have an off-topic doubt.
I’m using Sepharose IgG beads to purify a protein with a Protein A tag.
However, when I load fresh beads (without any protein) after incubating them
with 0.01% SDS or heating them at 95 °C in the presence of SDS, I observe a
strange band pattern on the SDS-PAGE
* With only 0.01% SDS: I see two consecutive bands between 72 and 95 kDa of
same intensity
* With heating: I see the same two bands (same intensity as above), plus
two strong bands at ~55 kDa and ~25 kDa .
The 55 kDa and 25 kDa bands are clearly the heavy and light chains, but I can’t
figure out what the upper bands are. I was wondering if anyone could help me
understand this.
Best regards,
Preeti
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