Anti-PKC (phospho S729) antibody [EPR1483(N)] - BSA and Azide free

• WB

Supplier Data

Western blot - Anti-PKC (phospho S729) antibody [EPR1483(N)] - BSA and Azide free (AB250360)

This data was developed using ab181119, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-PKC (phospho S729) antibody [EPR1483(N)] (<a href='/en-us/products/primary-antibodies/pkc-phospho-s729-antibody-epr1483n-ab181119'>ab181119</a>) at 1/2000 dilution

Lane 1:

Lysate of SH-SY5Y cells treated with Okadaic Acid and Calyculin A at 10 µg

Lane 2:

Lysate of SH-SY5Y cells at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated antibody at 1/1000 dilution

Predicted band size: 77 kDa

false

• WB

Unknown

Western blot - Anti-PKC (phospho S729) antibody [EPR1483(N)] - BSA and Azide free (AB250360)

This data was developed using ab181119, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PKC (phospho S729) antibody [EPR1483(N)] (<a href='/en-us/products/primary-antibodies/pkc-phospho-s729-antibody-epr1483n-ab181119'>ab181119</a>) at 1/2000 dilution

Lane 1:

THP-1 (human monocytic leukemia cell line) whole cell lysate at 15 µg

Lane 2:

THP-1 whole cell lysate treated with Phorbol-12-myristate-13-acetate at 100ng/ml for 24 hours at 15 µg

Lane 3:

THP-1 whole cell lysate treated with Phorbol-12-myristate-13-acetate at 100ng/ml for 24 hours. Then the membrane was incubated with phosphatase 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

Predicted band size: 63 kDa,77 kDa

Observed band size: 84 kDa

false

Exposure time: 3s

• WB

Unknown

Western blot - Anti-PKC (phospho S729) antibody [EPR1483(N)] - BSA and Azide free (AB250360)

This data was developed using ab181119, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : Lanes 1, 3, 5-8 : 10 seconds; Lanes 2, 4 : 3 minutes.

All lanes:

Western blot - Anti-PKC (phospho S729) antibody [EPR1483(N)] (<a href='/en-us/products/primary-antibodies/pkc-phospho-s729-antibody-epr1483n-ab181119'>ab181119</a>) at 1/1000 dilution

Lane 1:

Active human PKC alpha full length protein (<a href='/en-us/products/proteins-peptides/recombinant-human-pkc-alpha-protein-active-ab55672'>ab55672</a>) contains aa1-672 with GST-tag at 0.02 µg

Lane 2:

Active human PKC beta 1 full length protein (<a href='/en-us/products/proteins-peptides/recombinant-human-pkc-beta-1-protein-ab60840'>ab60840</a>) contains aa1-671 with GST-tag at 0.06 µg

Lane 3:

Active human PKC beta 2 full length protein (<a href='/en-us/products/proteins-peptides/recombinant-human-pkc-beta-2-protein-ab60841'>ab60841</a>) contains aa1-673 with GST-tag at 0.02 µg

Lane 4:

Active human PKC delta full length protein (<a href='/en-us/products/proteins-peptides/recombinant-human-pkc-delta-protein-ab60844'>ab60844</a>) contains aa1-676 with GST-tag at 0.06 µg

Lane 5:

Active human PKC eta full length protein (<a href='/en-us/products/proteins-peptides/recombinant-human-pkc-eta-protein-ab60849'>ab60849</a>) contains aa1-683 with GST-tag at 0.02 µg

Lane 6:

Active human PKC epsilon full length protein (<a href='/en-us/products/proteins-peptides/recombinant-human-pkc-epsilon-protein-ab60847'>ab60847</a>) contains aa1-737 with GST-tag at 0.02 µg

Lane 7:

Active human PKC theta full length protein (<a href='/en-us/products/proteins-peptides/recombinant-human-pkc-theta-prkcq-protein-ab56641'>ab56641</a>) contains aa1-706 with GST-tag at 0.02 µg

Lane 8:

Active human PKC mu full length protein (<a href='/en-us/products/proteins-peptides/recombinant-human-pkc-mu-pkd-protein-ab60873'>ab60873</a>) contains aa1-912 with GST-tag at 0.02 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

Predicted band size: 77 kDa

Observed band size: 100-150 kDa

false

• Dot

Unknown

Dot Blot - Anti-PKC (phospho S729) antibody [EPR1483(N)] - BSA and Azide free (AB250360)

This data was developed using ab181119, the same antibody clone in a different buffer formulation.

Dot blot analysis of PKC epsilon (pS729) phospho peptide (Lane 1), PKC epsilon non-phospho peptide (Lane 2), labelling PKC (pS729) with purified ab181119 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.