Anti-Tuberin (phospho S664) antibody [EPR8202]

• WB

Lab

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] (AB133465)

Blocking/Diluting buffer and concentration 5% NFDM/TBST Exposure time : Lane 1, 2 : 180 seconds Lane 3, 4 : 40 seconds Lane 5, 6 : 20 seconds

Lanes 1 - 6:

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] (ab133465) at 1/1000 dilution

Lanes 1 - 6:

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/tuberin-phospho-s664-antibody-epr8202-bsa-and-azide-free-ab248510'>ab248510</a>)

Lane 1:

Human brain lysate unboiled at 15 µg

Lane 2:

Human brain lysate unboiled, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

Lane 3:

Rat brain lysate unboiled at 15 µg

Lane 4:

Rat brain lysate unboiled, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

Lane 5:

Mouse brain lysate unboiled at 15 µg

Lane 6:

Mouse brain lysate unboiled, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 200 kDa

false

• WB

Lab

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] (AB133465)

Blocking/Diluting buffer and concentration 5% NFDM/TBST

Lanes 1 - 3:

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] (ab133465) at 1/1000 dilution

Lanes 1 - 3:

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/tuberin-phospho-s664-antibody-epr8202-bsa-and-azide-free-ab248510'>ab248510</a>)

Lane 1:

Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate unboiled at 15 µg

Lane 2:

HeLa treated with 200nM Phorbol-12-myristate-13-acetate (PMA) for 4 hours whole cell lysate unboiled at 15 µg

Lane 3:

HeLa treated with 200nM Phorbol-12-myristate-13-acetate (PMA) for 4 hours whole cell lysate unboiled, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 200 kDa

false

Exposure time: 80s

• WB

Unknown

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] (AB133465)

All lanes:

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] (ab133465) at 1/1000 dilution

Lane 1:

SH SY5Y cell lysates at 10 µg

Lane 2:

SH SY5Y cell lysates treated with Okadaic acid and Calyculin A at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 201 kDa

false

• WB

Lab

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] (AB133465)

Blocking/Diluting buffer and concentration 5% NFDM/TBST We recommend not to boil the samples after lysis to get desired WB results. The molecular weight observed is consistent with what has been described in the literature (PMID : 30774414; 10335945).

Lanes 1 - 4:

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] (ab133465) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/tuberin-phospho-s664-antibody-epr8202-bsa-and-azide-free-ab248510'>ab248510</a>)

Lane 1:

Mouse brain lysate boiled at 15 µg

Lane 2:

Mouse brain lysate unboiled at 15 µg

Lane 3:

Rat brain lysate boiled at 15 µg

Lane 4:

Rat brain lysate unboiled at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 200 kDa

false

Exposure time: 40s

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Tuberin (phospho S664) antibody [EPR8202] (AB133465)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.