Williams, who has won the event six times, was defeated at the Arthur Ashe Stadium by Australia’s Ajla TomljanoviÄ in three sets on Friday night.
The superstar broke down in tears as she thanked her mother, Oracene Price, and father, Richard Williams, for helping her in her legendary 25-year career.
“Oh my God, thank you so much, I tried but she played a bit better. Thank you daddy I know you are watching, thanks mom,” said an emotional Williams after the match.
“It all started with my parents, they deserve everything and I am really grateful for them. These are happy tears I guess, I don’t know. I wouldn’t be Serena unless there was Venus, so thank you, Venus.”
And she added: “It’s been a fun ride.”
Williams refused to fully commit to her retirement plans when asked if she would consider playing again.
“I don’t think so but you never know, I don’t know,” she said.
Asked after the match how she was feeling, TomljanoviÄ apologised to the crowd.
“I am feeling really sorry as I love Serena just as much as you do. This is a surreal moment for me,” she said.
Williams’s last ever singles loss came after wins over Anett Kontaveit of Estonia and Danka Kovinic of Montenegro in the first two rounds of her farewell tour.
The 40-year-old tennis legend has won 23 Major titles over her illustrious career, one short of the record held by Margaret Court and has said she will “evolve away from tennis” after a 27-year professional career.
She has not won a US Open championship since 2014, her third in a row. But her 367 Major match wins are the most by a female player in the open era.
The match was played in front of sister Venus and Williams’s family, as well as musician Seal, NFL player Russell Wilson and his wife, singer Ciara.
Congrats on an amazing career, @SerenaWilliams!
How lucky were we to be able to watch a young girl from Compton grow up to become one of the greatest athletes of all time.
I'm proud of you, my friend—and I can't wait to see the lives you continue to transform with your talents. pic.twitter.com/VWONEMAwz3
— Michelle Obama (@MichelleObama) September 3, 2022
Serena, THANK YOU. It is because of you I believe in this dream. The impact you’ve had on me goes beyond any words that can be put together and for that I say thank you, thank you, thank you, GOAT! pic.twitter.com/qeNZlC05WJ
— Coco Gauff (@CocoGauff) September 3, 2022
Her husband, Alexis Ohanian, was in the stands wearing a t-shirt with a photo of their daughter, Olympia, on the front.
The youngster was at her mum’s first- and second-round matches at the US Open, but was absent on Friday night.
I love you @serenawilliams It’s been the pleasure of a lifetime to watch you become what you have. Can’t wait to see what you do next. Thank you my old friend
— andyroddick (@andyroddick) September 3, 2022
.@serenawilliams you’re literally the greatest on and off the court. Thank you for inspiring all of us to pursue our dreams. I love you little sis!!!!!!
— Tiger Woods (@TigerWoods) September 3, 2022
— Oprah Winfrey (@Oprah) September 3, 2022
THANKS @serenawilliams YOU’RE GOING OUT SCRAPPING, JUST LIKE YOU CAME IN👊🏾👊🏾👊🏾‼️‼️‼️💯
— Samuel L. Jackson (@SamuelLJackson) September 3, 2022
The 40-year-old from Compton, California, broke TomljanoviÄ in the first set to take a 5-3 lead, before her opponent broke straight back.
TomljanoviÄ, 29, then broke Serena for a second time and served out to take the opening set 7-5 in 50 minutes with four consecutive games.
Williams then broke to open the second set, energizing the crowd and took a 4-0 lead before her opponent made a ferocious comeback.
Williams, roared on by a packed house, had multiple set points in an energy-sapping 15-minute eighth game of the set, but could not find a big shot to put it away.
She then had an opportunity to serve for the second set but TomljanoviÄ clawed her way back with a break of her own, before leveling it at 5-5.
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— Roland-Garros (@rolandgarros) September 3, 2022
— Wimbledon (@Wimbledon) September 3, 2022
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The set eventually went to a tiebreak, which Williams held her nerve to take and force a decisive third set.
Williams secured an early break in the final set but was then broken three times in a row by TomljanoviÄ who took a commanding lead and hushed the stunned crowd.
The fans gave Williams a standing ovation as her opponent served for the match and she bravely fought off five match points before TomljanoviÄ finally took the victory.
Former first lady Michelle Obama led tributes to the tennis legend, tweeting: “Congrats on an amazing career, @SerenaWilliams! How lucky were we to be able to watch a young girl from Compton grow up to become one of the greatest athletes of all time. I'm proud of you, my friend--and I can't wait to see the lives you continue to transform with your talents.”
Congratulations, Serena for an inspiring career! Cookie & I are so excited for you. I remember when we talked about you becoming an entrepreneur. Now, I look forward to watching you grow in business! @serenawilliams #Serena
— Earvin Magic Johnson (@MagicJohnson) September 3, 2022
What a run. @serenawilliams will be leaving behind an extraordinary tennis career that challenged the double standard between male and female players. I look forward to watching her continue building her amazing legacy off the court. https://t.co/K9Hxa2jB3I
— Bill Gates (@BillGates) September 3, 2022