Anti-KDEL antibody [EPR12668]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDEL antibody [EPR12668] (AB176333)

Immunohistochemical analysis of paraffin-embedded human liver sections labelling KDEL with purified ab176333 at dilution of 1 : 350. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. 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.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-KDEL antibody [EPR12668] (AB176333)

Overlay histogram showing 4% paraformaldehyde fixed HeLa cells labelling KDEL (red) with purified ab176333 at dilution of 1/100. The secondary antibody used was Alexa Fluor^® 488 goat-anti-rabbit IgG at dilution of 1/2000. A non-specific IgG antibody (rabbit monoclonal) was used as isotype control (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-KDEL antibody [EPR12668] (AB176333)

Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling KDEL with purified ab176333 at 1/200. Cells were fixed with 4% Paraformaldehyde and permeabilized using 0.1% Triton X-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/1000) using ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary. Nuclei were counterstained with DAPI (blue).

For negative control 1, ab150120 (Rabbit primary antibody) and anti-mouse secondary antibody were used and for negative control 2, ab7291 (Mouse primary antibody) and ab150077 (anti-rabbit secondary antibody) were used.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDEL antibody [EPR12668] (AB176333)

Immunohistochemical analysis of paraffin-embedded mouse liver sections labelling KDEL with purified ab176333 at dilution of 1 : 350. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. 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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDEL antibody [EPR12668] (AB176333)

Immunohistochemical analysis of paraffin-embedded rat kidney sections labelling KDEL with purified ab176333 at dilution of 1 : 350. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. 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.

• WB

Supplier Data

Western blot - Anti-KDEL antibody [EPR12668] (AB176333)

All lanes:

Western blot - Anti-KDEL antibody [EPR12668] (ab176333) at 1/1000 dilution

Lane 1:

293T cell lysate at 10 µg

Lane 2:

MCF7 cell lysate at 10 µg

Lane 3:

Raji cell lysate at 10 µg

Lane 4:

HepG2 cell lysate at 10 µg

Lane 5:

HeLa cell lysate at 10 µg

Lane 6:

Jurkat cell lysate at 10 µg

Lane 7:

K562 cell lysate at 10 µg

Predicted band size: 25 kDa

false

• WB

Lab

Western blot - Anti-KDEL antibody [EPR12668] (AB176333)

ab176333 detected several bands which have similar MWs with some proteins containing carboxyl terminal KDEL motifs, including P4HB, GRP78 BiP, GRP94, PDIA6 and LEPRE1 (PMID : 25683117, PMID : 19741001, PMID : 22079671, PMID : 28648146, PMID : 22615817). It indicates that ab176333 might recognize these proteins.

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

All lanes:

Western blot - Anti-KDEL antibody [EPR12668] (ab176333) at 1/5000 dilution

All lanes:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution

Predicted band size: 25 kDa

false

Exposure time: 3min

• WB

Unknown

Western blot - Anti-KDEL antibody [EPR12668] (AB176333)

Observed band size : 94,78,57,48

Blocking/Diluting buffer 5% NFDM/TBST

All lanes:

Western blot - Anti-KDEL antibody [EPR12668] (ab176333) at 1/10000 dilution

Lane 1:

Raw264.7 whole cell lysate at 20 µg

Lane 2:

C6 whole cell lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 25 kDa

false