Anti-CaMKII alpha (phospho T286) antibody [EPR1830] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CaMKII alpha (phospho T286) antibody [EPR1830] - BSA and Azide free (AB248035)

This data was developed using ab124880, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of Paraffin-embedded sections Mouse Cerebellum tissue labelling CaMKII (pT286) with ab124880 at 1/5000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution. Mouse cerebellum without Lambda Protein Phosphatase treatment staining on Mouse Cerebellum is observed. Counterstained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution.

Positive staining on mouse cerebellum without Lambda Protein Phosphatase treatment (image A); No signal was detected when tissues were treated with Lambda Protein Phosphatase (image B).

The section was incubated with ab124880 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CaMKII alpha (phospho T286) antibody [EPR1830] - BSA and Azide free (AB248035)

Immunohistochemical analysis of Paraffin-embedded sections Rat Cerebellum tissue labelling CaMKII (pT286) with ab124880 at 1/5000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)(Ready to use). Rat cerebellum without Lambda Protein Phosphatase treatment staining on Rat Cerebellum is observed. Counterstained with Hematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)(Ready to use). Positive staining on rat cerebellum without Lambda Protein Phosphatase treatment (image A); No signal was detected when tissues were treated with Lambda Protein Phosphatase (image B). The section was incubated with ab124880 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins. This data was developed using ab124880, the same antibody clone in a different buffer formulation.

• WB

Lab

Western blot - Anti-CaMKII alpha (phospho T286) antibody [EPR1830] - BSA and Azide free (AB248035)

Blocking and diluting buffer and concentration : 5% NFDM/TBST This data was developed using ab124880, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-CaMKII alpha (phospho T286) antibody [EPR1830] (<a href='/ko/products/primary-antibodies/camkii-alpha-phospho-t286-antibody-epr1830-ab124880'>ab124880</a>) at 1/4000 dilution

Lane 1:

Rat brain tissue lysate (untreated membrane) at 20 µg

Lane 2:

Rat brain tissue lysate (Lambda phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 50 kDa,54 kDa

false

Exposure time: 92s

• WB

Lab

Western blot - Anti-CaMKII alpha (phospho T286) antibody [EPR1830] - BSA and Azide free (AB248035)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. This data was developed using ab124880, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-CaMKII alpha (phospho T286) antibody [EPR1830] (<a href='/ko/products/primary-antibodies/camkii-alpha-phospho-t286-antibody-epr1830-ab124880'>ab124880</a>) at 1/4000 dilution

Lane 1:

Mouse brain tissue lysate (untreated membrane) at 20 µg

Lane 2:

Mouse brain tissue lysate (Lambda phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 54 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-CaMKII alpha + CaMKII beta (phospho T286) antibody [EPR1830] - BSA and Azide free (AB248035)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. This data was developed using ab248035, the same antibody clone in a different buffer formulation.

Lane 1:

Anti-LMAN2 antibody (ab213529) at 0.25 µg/mL

Lanes 1 - 2:

Western blot - Anti-CaMKII alpha (phospho T286) antibody [EPR1830] (<a href='/ko/products/primary-antibodies/camkii-alpha-phospho-t286-antibody-epr1830-ab124880'>ab124880</a>) at 1/4000 dilution

Lanes 1 - 2:

Western blot - Anti-CaMKII alpha + CaMKII beta (phospho T286) antibody [EPR1830] - BSA and Azide free (ab248035) at 1/4000 dilution

Lanes 2 - 3:

Anti-LMAN2 antibody (ab213529) at 0.5 µg/mL

Lanes 1 - 2:

Whole extract of HEK-293T cells overexpressing Human LMAN2

Lane 1:

Mouse brain tissue lysate (untreated membrane) at 20 µg

Lane 2:

Mouse brain tissue lysate (Lambda phosphatase treated membrane) at 20 µg

Lane 3:

Whole extract of HEK-293T cells overexpressing Human LMAN2 with LMAN2 immunizing peptide

Secondary

Lanes 1 - 3:

Goat Anti-Rabbit IgG-Peroxidase

Lanes 1 - 2:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-CaMKII alpha (phospho T286) antibody [EPR1830] - BSA and Azide free (AB248035)

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.