一般社団法人
日本細胞生物学会Japan Society for Cell Biology

染めている人からの情報
Information from other users
繊毛が染まるはず
Expected to stain cilia

3%PFA (in PBS)

3%PFA (in PBS)

【PFA+Triton】:○,【PFA+Saponin】:○【PFA条件】3%PFA/PBS　37℃10min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS

3%PFA in PBS

0.1%Triton

線毛を見るには、1次抗体は1%NGS, 0.1%Triton　4度overnightで染色する必要あり！！（0.05% saponinだと細胞内の微小管が多く染まるため不適）
To visualize cilia, use 1st Ab in 1%NGS, 0.1%Triton overnight 4C, BG very high by 0.05% saponin.

【PFA+Triton】:○,【PFA+Saponin】:○【PFA条件】3%PFA/PBS　37℃10min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS

4%PFA, MeOH, TCA

4% PFA

For apoptosis detection 4%PFA, TCA

4% PFA at 4oC over night

0.5% Triton in PBS

4%PFA (in phosphate buffer)

3%PFA (in PBS)

0.05%saponin

3%PFA (in phosphate buffer)

3%PFA (in PBS)

0.05%saponin

3%PFA (in phosphate buffer)

3%PFA (in PBS)

0.05%saponin

3%PFA (in phosphate buffer)

3%PFA (in PBS)

0.05%saponin

fine punctate staining

3%PFA (in PBS)

0.05%saponin

fine punctate staining

3%PFA (in PBS)

0.05%saponin

fine punctate staining (similar to alpha)

3%PFA (in PBS)

0.05%saponin

fine punctate staining (similar to alpha)

3%PFA (in PBS)

0.05%saponin

fine punctate staining

3%PFA (in PBS)

0.05%saponin

fine punctate staining (larger than alpha)

3%PFA (in PBS)

0.05%saponin

fine punctate staining (larger than alpha)

【PFA+Triton】: -OA:○, +OA:×,【PFA+Saponin】: -OA:○, +OA:△,【MeOH 】:×,【PFA+70%EtOH】: -OA:○, +OA:△【PFA条件】3%PFA/PBS　37℃10min【MeOH条件】MeOH 100% -20℃ 5min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS

MeOH

WB:Blocking/3%BSA-TBS, 1stAb/3%BSA-TBS, 2ndAb/3% skin milk-TBSが良い。

4%PFA (in PBS)

PFA-fixed cryosection/HistoVT one (nacalai) 70˚C 20min

astrocyte

ELISA用

4 % PFA fix, PBS+0.05 %Tween20+ 0.5 % TSA block+1 % Donkey serum

3%PFA (in PBS)

0.05%saponin

apical staining

3%PFA (in PBS)

0.05%saponin

MeOH, TCAでストレスファイバー弱く染める
Stress fibers are stained by MeOH or TCA fix.

TCA

3%PFA (in PBS)

4% PFA at 4oC over night

0.5% Triton in PBS

3%PFA (in PBS)

0.05%saponin

included in Coated vesicle sampler kit

3%PFA (in phosphate buffer)

IF: -20℃ methanol or acetone:methanol=1:1

Can Get Signal (TOYOBO)の使用が望ましい Favorable to use Can Get Signal (TOYOBO)

IF: -20℃ methanol or acetone:methanol=1:1

3%PFA (in PBS)

0.05%saponin

3%PFA (in PBS)

0.05%saponin

4％PFA +0.1％Triton

4% PFA

100%EtOH/Acetone

【PFA+Triton】:×,【IS】:○？【PFA条件】3%PFA/PBS　37℃10min【Triton条件】0.1%　Triton-X100/PBS

【PFA+Triton】:×,【IS】:○？【PFA条件】3%PFA/PBS　37℃10min【Triton条件】0.1%　Triton-X100/PBS

【PFA+Triton】:×,【IS】:○？,【TCA】:○【PFA条件】3%PFA/PBS　37℃10min【TCA条件】TCA 10%/H2O on ice 15min【Triton条件】0.1%　Triton-X100/PBS

IFはTCA固定
TCA fix is best for IF.

【PFA+Triton】:○,【PFA+Saponin】:○【PFA条件】3%PFA/PBS　37℃10min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS

WB:バンドが二本
Two bands appear by WB.

【PFA+Triton】:○,【PFA+Saponin】:○【PFA条件】3%PFA/PBS　37℃10min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS

WB:バンドが一本、お勧め
A unique band appear by WB. Recommended Ab.

【PFA+Triton】:◎【PFA条件】3%PFA/PBS　37℃10min【Triton条件】0.1%　Triton-X100/PBS

【PFA+Triton】:○,【PFA+Saponin】:○【PFA条件】3%PFA/PBS　37℃10min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS

4%PFA (in phosphate buffer)

4％PFA +0.1％Triton

3%PFA (in PBS), methanol

1%PFA 染まるが核染が強い。
Nonspecific nuclear staining.

MeOH fixation

0.05% triton X-100

mouse specific

WBのみ使用可能

【PFA+Triton】:×,【PFA+Saponin】:×,【MeOH 】:×,【PFA+70%EtOH 】:×,【PFA_digitonin】:△(Can Get B)【PFA条件】3%PFA/PBS　37℃10min【MeOH条件】MeOH 100% -20℃ 5min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS【digitonin条件】50ug/ml digitonin /PBS

ice cold Methanol

ice cold Methanol

ice cold Methanol

TCA, 4%PFA, 1%PFA, MeOH

3%PFA (in PBS)

4％PFA +0.1％Triton

3%PFA (in PBS)

マウス小腸ではこれがベスト
Best Ab of all alkaline phosphatase Abs for frozen section of mouse small intestine.

3%PFA (in PBS)

【PFA+Triton】:◎,【PFA+Saponin】:○【PFA条件】3%PFA/PBS　37℃10min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS

MDCKII OK。Triton, mouse488がいい。
Alexa488 anti-mouse antibody is suitable for 2nd Ab.

【PFA+Triton】:○,【PFA+Saponin】:◎【PFA条件】3%PFA/PBS　37℃10min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS

saponin, mouse555がいい。
Better to use saponin and Alexa555 2nd antibody.

3%PFA (in PBS)

IFではよくわからないが、ImEMではシナプス小胞が染まっているように見える
By ImEM, synaptic vesicles can be stained by this Ab, but staining is not evident by IF.

3%PFA (in PBS)

3%PFA (in PBS)

3%PFA (in PBS), methanol

3%PFA (in PBS)

3%PFA (in PBS), methanol

1%PFA, 4%PFA, MeOH, TCA

試してない？
Not used

3%PFA (in PBS)

3%PFA (in PBS)

3%PFA (in PBS)

3%PFA (in PBS)

3%PFA (in PBS)パパインの前処理必要
Pretreatment by papain is required

ＯＫ（北大渡辺先生作）パパインによる前処理必要
Made by Prof M. Watanabe (Hokkaido Univ.) pretreatment by papain is required.

【4%PFA+0.2%Saponin】:○

【Formalin in PBS+0.05%Triton】:○

ホルマリンは37％ホルマリン液（ナカライ）をPBSやRingerで１/10に希釈
Formalin solution is made by dilution of 37% formalin (Nacalai Inc.) by PBS or Ringer's solution

WB:Blocking/3%BSA-TBS, 1stAb/3%BSA-TBS, 2ndAb/3% skin milk-TBSが良い。

4%PFA/PBS

ice cold Methanol

4%PFA/PBS

ice cold Methanol

ice cold Methanol

ice cold Methanol

ice cold Methanol

ice cold Methanol

ice cold Methanol

4%PFA/PBS

3%PFA (in PBS)

0.05%saponin

fine punctate staining like adaptin alpha , included in Coated vesicle sampler kit

3%PFA (in PBS), methanol

不明瞭

3%PFA (in PBS)

4% PFA at 4oC over night

0.5% Triton in PBS

MeOH fixation

0.05% triton X-100

MeOH fixation

0.05% triton X-100

MeOH, TCA

MeOH, TCA

methanol 固定可能
methanol fixation also applicable

【抗原の種、部位】N-terminal peptide

WBに使わないほうがいい、IFもSA4の方がきれい。
Not suitable for WB. For IF, antibody SA4 is better than this antibody.

PFA+Triton or PFA/GA+Triton (fix cells by adding 1/10 vol of 37% formalin directly to the culture medium)

PFAは37％ホルマリン液を培地に1/10直接加えて固定する fix cells by adding 1/10 vol of 37% formalin directly to the culture medium

4％PFA +0.1％Triton

4％PFA +0.1％Triton

3%PFA (in PBS)

0.05%saponin

3%PFA in PBS

0.05% saponin

3%PFA (in PBS)

0.05%saponin

PFA (fix cells by adding 1/10 vol of 37% formalin directly to the culture medium)+Triton or MeOH

PFA (fix cells by adding 1/10 vol of 37% formalin directly to the culture medium)+Triton or MeOH

4%PFA (in PBS)

HCl, RT, 30m

4%PFA (in PBS)

HCl, RT, 30m

4％PFA +0.1％Triton

3%PFA (in PBS)

特殊な前処理が必要（普通に染めたら染まらないので注意）
Pretreatment should be required unless no signal.

3%PFA (in PBS)

3%PFA (in PBS)

3%PFA (in PBS)

4% PFA at 4oC over night

0.5% Triton in PBS

3%PFA

抗原の賦活化が必要らしい
Antigen retreival should be required

【PFA+Triton】:○【PFA条件】3%PFA/PBS　37℃10min【Triton条件】0.1%　Triton-X100/PBS

IFの時、0.1% Triton+0.05%SDSを使う。
For IF, use 0.1% Triton+0.05%SDS

Ptk2 ダメ、MDCKII OK。
Not suitable for PtK2.

4%PFA, 1%PFA, MeOH, TCA

4%PFA (in PBS)

PFA-fixed cryosection/HistoVT one (nacalai) 70˚C 20min

mature DG granule cell, adult CA1 pyramidal cell, a type of interneuron

4%PFA (in PBS)

PFA-fixed cryosection/HistoVT one (nacalai) 70˚C 20min

mature DG granule cell, adult CA1 pyramidal cell, a type of interneuron

4%PFA (in PBS)

PFA-fixed cryosection/HistoVT one (nacalai) 70˚C 20min

mature DG granule cell, adult CA1 pyramidal cell, a type of interneuron

3%PFA (in PBS)

3%PFA (in phosphate buffer)

pretreatment with 5%normal donkey serum, 0.1% TritonX-100 in PBS RT for 1hr before 1st antibody is recommended

【PFA+Triton】:×,【MeOH】:△,【TCA】:×【PFA条件】3%PFA/PBS　37℃10min【MeOH条件】MeOH 100% -20℃ 5min【TCA条件】TCA 10%/H2O on ice 15min【Triton条件】0.1%　Triton-X100/PBS

【4%PFA+0.2%Saponin】:○

3%PFA (in PBS)

0.05%saponin

3%PFA in PBS

0.05% saponin

Ab in Chaperone sampler kit (BD Transduction Lab)

3%PFA in PBS

0.05% saponin

Ab in Chaperone sampler kit (BD Transduction Lab)

3%PFA (in PBS)

0.05%saponin

殆どの細胞で核膜は染まる。染まる細胞と染まらない細胞の差が大きい
Nuclear envelopes are stained in most cells. High variation in staining intensities among cells.

3%PFA (in PBS)

0.05%saponin

3.7% Formaldehyde (in PBS)

Methanol (-20˚C)

methanol

3%PFA in PBSでも染まるが弱い
Only weakly stained by 3%PFA in PBS fixation

4%PFA (in PBS)

4%PFA (in PBS)

PFA-fixed cryosection/HistoVT one (nacalai) 70˚C 20min

immature DG granule cell, a type of interneuron, Cajal-retzius cell

4%PFA (in PBS)

PFA-fixed cryosection/HistoVT one (nacalai) 70˚C 20min

immature DG granule cell, a type of interneuron, Cajal-retzius cell

4%PFA (in PBS)

PFA-fixed cryosection/HistoVT one (nacalai) 70˚C 20min

4 % PFA fix, PBS+0.05 %Tween20+ 0.5 % TSA block+1 % Donkey serum

【PFA+Triton】:×,【PFA+Saponin】:◎,【MeOH】:×【PFA条件】3%PFA/PBS　37℃10min【MeOH条件】MeOH 100% -20℃ 5min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS

Lamp1, LBPAと共局在する。
Colocalize with lamp1 and LBPA.

【PFA+Triton】:×,【PFA+Saponin】:◎,【MeOH】:×【PFA条件】3%PFA/PBS　37℃10min【MeOH条件】MeOH 100% -20℃ 5min【Triton条件】0.1%　Triton-X100/PBS【saponin条件】0.05%　saponin /PBS

Lamp1, LBPAと共局在する。
Colocalize with lamp1 and LBPA.

3%PFA (in PBS)

0.05%saponin

【PFA+Saponin】:○【PFA条件】3%PFA/PBS　37℃10min【saponin条件】0.05%　saponin /PBS

染まり方にムラがある。
Uneven staining

【3.7% formalin solution +0.5%Triton】:○ formalin solution is made by dilution of 37% formalin (Nacalai Inc.) by PBS or Ringer's solution

ホルマリンは37％ホルマリン液（ナカライ）をPBSやRingerで１/10に希釈
Formalin solution is made by dilution of 37% formalin (Nacalai Inc.) by PBS or Ringer's solution