Campaigners from Rape Crisis Scotland (RCS) said the hard-hitting advert was specifically produced to make people "stop in their tracks" and challenge "women-blaming" beliefs that dressing provocatively, being drunk or flirting with men are contributory factors.
The advert, which was due to be aired at 9:30pm during the Brazil versus Chile game, shows a young woman wearing a short blue skirt enjoying herself in a bar.
A man says to a friend: "Check out the skirt, she's asking for it."
The "Not Ever" advert then cuts to earlier in the day when the woman tells a shop assistant, "I'm going out tonight and I want to get raped. I need a skirt that'll encourage a guy to have sex with me against my will."
She then looks at the camera and says in a voice laden with irony: "As if". The voiceover continues: "Nobody asks to be raped. Not ever."
Sandy Brindley, national co-ordinator of RCS, said: "We know this advert will make people stop in their tracks but that's the intention. Recent research reveals that almost one in five Scots believe a woman is partially to blame for being raped if she is wearing revealing clothing, which is a ridiculously unjust and insensitive judgement to make.
"The advert has been designed to shake out ingrained prejudices many Scots have towards women who have been raped, in a bid to stop perpetuating the myths surrounding rape. Even though people believe they wouldn't judge a rape victim by what they wear, how drunk they were or if they had been flirting, they often actually do."
Ms Brindley added: "This campaign aims to prompt people to keep their judgements in check and to remember that there's only one person who is responsible for rape, and it's not the victim. It doesn't matter what you wear, how many sexual partners you've had or if you're out drinking with friends - no-one deserves to be raped, not ever."
Fay Wilson, a rape survivor who waived her anonymity earlier this year to feature in the first phase of the RCS campaign "This is not an invitation to rape me", said: "I do think this advert will reach the target audience and really get through to them. The fact it's being aired during the World Cup means a lot of people will be watching and that will get people talking about it and thinking about these things."
But Bill Aitken MSP, Tory justice spokesman, said: "I'm not too sure this advert is as clear as it should be. While the intentions are obviously good, some viewers might think the woman was not speaking ironically and that the man's verbal behaviour was acceptable.
"I do think the message should have been much more focused."
The Scottish Government-funded 30-second advert cost a total of 160,000 including production costs and a seven-week run on Scottish Television, Channel 4 and Channel 5.
Hollywood actress Laura Fraser, who has starred in hits including A Knight's Tale and The Man in the Iron Mask, said: "I support this campaign because I feel we need to understand that our perspective regarding rape is warped. I think if we imagine our sister, daughter or mother in this scenario then things will look different.
"Changing these women-blaming attitudes is a solid starting point for making women safer in Scotland."
A Scottish Government spokesperson said: "This campaign will help stress that rape is never acceptable and challenge the dangerous misconception that victims are to blame." A survey of 1,040 Scots carried out by Cello MRUK in February 2010 for the Scottish Government found that:
• 23 per cent think a woman can be at least partly responsible if she is drunk at the time of the attack
• 17 per cent think that a woman bears some responsibility if she wears revealing clothing
• 15 per cent say there should be some burden of responsibility for rape if the women is flirting
The campaign website can be viewed on www.notever.co.uk.