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人CC趨化因子受體(CCR1)試劑盒
  • 人CC趨化因子受體(CCR1)試劑盒
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更新時間:2024-12-10 21:00:07

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SIGMA現貨,SIGMA代理

人CC趨化因子受體(CCR1)試劑盒

人CC趨化因子受體(CCR1)試劑盒報價,咨詢選購,技術服務,選擇正確的實驗試劑或材料是實驗成功的關鍵。

  一、客戶收到細胞先不開蓋,放在培養箱靜置若干小時后(看細胞密度而定)在倒置顯微鏡下觀察細胞生長情況,并對細胞進行不同倍數拍照(建議受收細胞時的培養基拍一張照片,觀察培養基的顏色和是否有漏液情況,顯微鏡下拍細胞100X,200X各一張),排除細胞本身污染的情況;

  收到細胞未開封,出現污染狀況我們負責免費發送一株細胞。

  二、如為貼壁細胞,請在顯微鏡下觀察細胞密度:

  1. 未超過80%匯合度時,用75%酒精(建議配置75%酒精的水也是滅菌超純水,噴灑整個瓶消毒后放到超菌臺內,嚴格無菌操作;然后打開蓋子,先把培養基吸出10ml左右,放在離心管里,然后瓶口過火(嚴禁直接傾倒),這樣可以保證不污染,再用10ml的移液管,將剩余的培液全部吸出,放于離心管中,培養瓶中只剩余10ml左右培液就可以了。培養16hr(時間根據細胞生長情況調整)之后,傳代或者凍存。

  2. 超過80%匯合度時,請按細胞培養條件傳代培養。具體步驟如下:

  1) 棄去培養液,用3ml PBS(不含鈣,鎂離子)洗1-2次;

  2) 加1ml消化液(0.25%Trypsin-0.53mM EDTA)于T25培養瓶中,倒轉放于37度培養箱1-3分鐘預熱,然后又將培養瓶倒轉大約30秒后,在倒置顯微鏡下觀察細胞消化情況,若細胞大部分變圓分散,輕敲幾下培養培養瓶,細胞隨即脫落下來;

  3) 加入6-8ml*培養基,吸出,分到新的培養瓶中,按要求分瓶。

  三、如為懸浮細胞,吸出培養液,1000轉/分鐘離心5分鐘,吸出上清,管底細胞用新鮮培養基懸浮細胞后移回培養瓶。

  注意:換液時一半用我們的培養基,一半用你們的,以免細胞不適應而造成生長不好。

  四、細胞出現問題,可重發的情況有哪些?判定標準是什么?

  (1)細胞運輸途中遭遇的各種問題,細胞丟失、破損、漏液等,重發;

  (2)凍存的細胞復蘇后,絕大多數細胞未存活,重發;

  (3)存活的細胞靜置24小時后,絕大多數細胞未存活,重發;

  (4)凍存的細胞復蘇后或存活的細胞靜置4小時后并且未開封,出現污染,重發;

  (5)視具體情況而定。 

    細胞培養技術3個原則:1、用自己的一套試劑,不要和別人混用;2、勤觀察細胞,像養寵物一樣喜愛和關心;3、有規律地換液和傳代,不要“三天打魚,兩天曬網”。凍存細胞時間為5-7個工作日,活細胞為7-15個工作日。

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