The ALP candidate for the Queensland seat of Longman, Susan Lamb, won convincingly in the by-election, indicating there was grass roots support for the Adani project.
The Stop Adani movement said the ALP had not actually committed to stopping the mine if it won government.
It wants protestors to target Shorten, opposition treasury spokesman Chris Bowen and opposition spokesman for Resources and Northern Australia Jason Clare, and say: "I want Labor to commit to Stop Adani".
"We know that Adani's Carmichael coal mine doesn't stack up environmentally or economically," Stop Adani said.
"We need our politicians to know it won't stack up for them politically, either.
"Whatever happens with Adani's finance, we can't afford to wait. We need a political stop to end Adani's mine plans.
"Getting Labor to publicly commit to stop Adani's mine is critical. If Labor commits to stopping the mine, this will scare Adani's investors away. If enough people tell Labor MPs to get off the fence and commit to stopping this disastrous mine once and for all we'll be one step closer to stopping Adani."
They could have a bit of a problem with that though because a couple of key Labor players - the Construction Forestry Mining and Energy Union and the Australian Workers Union - both favour the mine, albeit conditionally.
The CFMEU is the most powerful mine in the Queensland coalfields and will have a big influence on Shorten's position.
"We do support the mine," CFMEU Mining Queensland district president Stephen Smyth told Australia's Mining Monthly.
"It must be economically viable and meet the required legislative requirements for environmental impact statement.
"It must meet the occupational health and safety requirements and the Native Title requirements and meet industry standard collective agreements for employment.
"Finally it must employ local workers in secure well-paying jobs.
"We do not support this mine opening up using a labour hire or contractual workforce in place of good well-paying secure employment. Also they must look locally for employees."
The other aspect is the union does not support the idea of the Adani mine using automation in the place of people.
"This is the same approach applies across all mining companies," Smyth said.
"We have a position that if any mining company wishes to use automation for equipment then they should lose the right to the diesel fuel rebate for starters."
The AWU - which Shorten once led - said coal mining was a major part of its workers' supply chain.
"We are a heavy intensive manufacturing union, for steel and aluminium, we rely on coal as base load power generation to keep most of these going," AWU secretary Daniel Walton told Sky News.
"Australia needs a coal industry going forward and if this project, like so many others, stacks up viably, financially and everything else, then it should go ahead."