Anti-CREB (phospho S133) antibody [E113] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB (phospho S133) antibody [E113] - BSA and Azide free (AB220798)

This data was developed using ab32096, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human astrocytoma tissue labeling CREB with ab32096 at 1/2000 followed by a ready to use ab209101. Nuclear staining on human astrocytoma without alkaline phosphatase treatment (image A). No signal was detected when tissues were treated with alkaline phosphatase (image B). The section was incubated with ab32096 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND®RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use ab209101. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CREB (phospho S133) antibody [E113] - BSA and Azide free (AB220798)

Intracellular Flow Cytometry analysis of A431 (human epidermoid carcinoma) cells labeling CREB with purified ab32096 at 1/70 dilution (10μg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32096).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB (phospho S133) antibody [E113] - BSA and Azide free (AB220798)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue labelling CREB with purified ab32096 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32096).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB (phospho S133) antibody [E113] - BSA and Azide free (AB220798)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid gland adenocarcinoma tissue labelling CREB with unpurified ab32096 at 1/250 dilution.

Panel A : Cells are untreated.
Panel B : Cells are treated with Phosphatase.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32096).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CREB (phospho S133) antibody [E113] - BSA and Azide free (AB220798)

Immunocytochemistry/Immunofluorescence analysis of LP treated and untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) labelling CREB with purified ab32096 at 1/200. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain at a dilution of 1/200.

Confocal image showing nuclear staining on HeLa cells .The signal decreased after Lambda Protein Phosphatase treatment ( 31?,2h).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32096).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CREB (phospho S133) antibody [E113] - BSA and Azide free (AB220798)

Immunocytochemistry/Immunofluorescence analysis of A431 cells labelling CREB with unpurified ab32096 at 1/250.

Panel A : Cells are untreated.

Panel B : Cells are treated with Phosphatase.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32096).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CREB (phospho S133) antibody [E113] - BSA and Azide free (AB220798)

Immunocytochemistry/Immunofluorescence analysis of A431(human epidermoid carcinoma) cells +/- AP 37? 1h labelling CREB with purified ab32096 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1 : primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).

Control 2 : ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32096).

• IP

Unknown

Immunoprecipitation - Anti-CREB (phospho S133) antibody [E113] - BSA and Azide free (AB220798)

ab32096 (purified) at 1/50 immunoprecipitating CREB (phospho S133) in HeLa whole cell lysate. 10 ug of cell lysate was present in the input. For western blotting, a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) was used for detection at 1/1,500 dilution. A rabbit monoclonal IgG (ab172730) was used intead of ab128913 as a negative control (Lane 3).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32096).

All lanes:

Immunoprecipitation - Anti-CREB (phospho S133) antibody [E113] (<a href='/en-us/products/primary-antibodies/creb-phospho-s133-antibody-e113-ab32096'>ab32096</a>)

Predicted band size: 36 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-CREB (phospho S133) antibody [E113] - BSA and Azide free (AB220798)

ab32096 at 1/100 dilution immunoprecipitating CREB (phospho S133) in HeLa (human cervix adenocarcinoma) treated with 25ug/mL anisomycin for 30 minutes, whole cell lysate, observed at 40 kDa (lanes 1 and 2).

Lane 1 (input) : HeLa treated with 25ug/mL anisomycin for 30 minutes. Whole cell lysate, 10μg.

Lane 2 (+) : ab32096 + HeLa treated with 25ug/mL anisomycin for 30 minutes. Whole cell lysate.

Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab32096 in treated with 25ug/mL anisomycin for 30 minutes. Whole cell lysate

For western blotting, ab32096 at 1/200 dilution followed by ab131366 VeriBlot for IP (HRP) at 1/1000 as the secondary antibody.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32096).

All lanes:

Immunoprecipitation - Anti-CREB (phospho S133) antibody [E113] (<a href='/en-us/products/primary-antibodies/creb-phospho-s133-antibody-e113-ab32096'>ab32096</a>)

Predicted band size: 36 kDa

false

Exposure time: 5s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB (phospho S133) antibody [E113] - BSA and Azide free (AB220798)

This data was developed using ab32096, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CREB with ab32096 at 1/2000 followed by a ready to use ab209101. Nuclear staining on mouse cerebrum without alkaline phosphatase treatment (image A). No signal was detected when tissues were treated with alkaline phosphatase (image B). The section was incubated with ab32096 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND®RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use ab209101. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB (phospho S133) antibody [E113] - BSA and Azide free (AB220798)

This data was developed using ab32096, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling CREB with ab32096 at 1/2000 followed by a ready to use ab209101. Nuclear staining on rat cerebrum without alkaline phosphatase treatment (image A). No signal was detected when tissues were treated with alkaline phosphatase (image B). The section was incubated with ab32096 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND®RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use ab209101. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

AbReview21176****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREB (phospho S133) antibody [E113] - BSA and Azide free (AB220798)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat hippocampus tissue labelling CREB with unpurified ab32096 at 1/100 dilution. Sections were subjected to antigen retrieval by autoclave prior to blocking with 8% milk for 30 minutes at 37°C. The primary antibody was diluted 1/100 with DAKO antibody diluent and incubated with the sample for 18 hours at 4°C. An LSAB-labeled Streptavidin-Biotin conjugated Goat polyclonal antibody was used undiluted as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32096).

This image is courtesy of an Abreview submitted by Akiko Shingo.

• Dot

Supplier Data

Dot Blot - Anti-CREB (phospho S133) antibody [E113] - BSA and Azide free (AB220798)

Dot blot analysis of CREB (pS133) phospho peptide (Lane 1) and CREB non-phospho peptide (Lane 2) using ab32096 at 1/1000 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution.

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

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 (ab32096).