Anti-JAK1 (phospho Y1034 + Y1035) antibody [EPR1899(2)] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK1 (phospho Y1034 + Y1035) antibody [EPR1899(2)] - BSA and Azide free (AB203784)

Immunohistochemical analysis of paraffin-embedded human liver carcinoma sections labelling JAK1 (phospho Y1034 + Y1035) with purified ab138005 at dilution of 1/50. The secondary antibody used was ImmunoHistoProbe HRP Polymer for Rabbit IgG at dilution of 1 : 0. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab138005).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK1 (phospho Y1034 + Y1035) antibody [EPR1899(2)] - BSA and Azide free (AB203784)

Immunohistochemical analysis of paraffin embedded Human colon adenocarcinoma tissue labelling JAK1 with unpurified ab138005 antibody at 1/50 dilution. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab138005). Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-JAK1 (phospho Y1034 + Y1035) antibody [EPR1899(2)] - BSA and Azide free (AB203784)

Blocking and diluting buffer 5% NFDM/TBST. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab138005).

All lanes:

Western blot - Anti-JAK1 (phospho Y1034 + Y1035) antibody [EPR1899(2)] (<a href='/en-us/products/primary-antibodies/jak1-phospho-y1034-y1035-antibody-epr18992-ab138005'>ab138005</a>) at 1/500 dilution

Lane 1:

Untreated Ramos whole cell lysate at 10 µg

Lane 2:

Ramos treated with 1mM pervanadate for 30 minutes whole cell lysate at 10 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: 133 kDa

Observed band size: 133 kDa

false

• ELISA

Lab

ELISA - Anti-JAK1 (phospho Y1034 + Y1035) antibody [EPR1899(2)] - BSA and Azide free (AB203784)

Antigen :

1. JAK1 (pY1034) phospho peptide
2. JAK1 (pY1035) phospho peptide
3. JAK1 non-phospho peptide

Antigen concentration : 0~1000ng/ml

Primary antibody concentration range : 1000ng/ml

Secondary antibody : Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), 1/2500 dilution

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab138005).

• WB

Lab

Western blot - Anti-JAK1 (phospho Y1034 + Y1035) antibody [EPR1899(2)] - BSA and Azide free (AB203784)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab138005).

Blocking and diluting buffer and concentration : 5% NFDM /TBST.

ab181602 was used as a GAPDH loading control.

Samples are non-boiled as boiling may cause protein aggregation.

All lanes:

Western blot - Anti-JAK1 (phospho Y1034 + Y1035) antibody [EPR1899(2)] (<a href='/en-us/products/primary-antibodies/jak1-phospho-y1034-y1035-antibody-epr18992-ab138005'>ab138005</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with a Human JAK1 expression vector containing a His tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a Human JAK1 expression vector containing a His tag were treated with 50uM pervanadate for 30 min whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a Human JAK1 (mutated Y1034A, mutated Y1035A) expression vector containing a His tag whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a Human JAK1 (mutated Y1034A, mutated Y1035A) expression vector containing a His tag were treated with 50uM pervanadate for 30 min whole cell lysate at 20 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: 133 kDa

Observed band size: 133 kDa

false

Exposure time: 180s

• Dot

Lab

Dot Blot - Anti-JAK1 (phospho Y1034 + Y1035) antibody [EPR1899(2)] - BSA and Azide free (AB203784)

Primary antibody : 1/1000 (unpurified)

Secondary antibody : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051)

Secondary ab dilution : 1/100,000

Blocking buffer and concentration : 5% NFDM/TBST

Diluting buffer and concentration : 5% NFDM /TBST

Lane 1 : JAK1 (pY1034) phospho peptide

Lane 2 : JAK1 (pY1035) phospho peptide

Lane 3 : JAK1 (pY1034/pY1035) phospho peptide

Lane 4 : JAK1 non-phospho peptide

Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab138005).

• OI-RD Scanning

Lab

OI-RD Scanning - Anti-JAK1 (phospho Y1034 + Y1035) antibody [EPR1899(2)] - BSA and Azide free (AB203784)

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.