Anti-ARPC2 antibody [EPR8533]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ARPC2 antibody [EPR8533] (AB133315)

Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling ARPC2 with purified ab133315 at 1/500 dilution. Cells were fixed with 100% methanol. ab150077 Goat anti rabbit IgG (Alexa Fluor^®488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARPC2 antibody [EPR8533] (AB133315)

Immunohistochemical analysis of paraffin embedded human tonsil tissue labelled with ab133315 (unpurified) at 1/250 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARPC2 antibody [EPR8533] (AB133315)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric carcinoma tissue sections labeling ARPC2 with purified ab133315 at 1/2000 dilution (0.25 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ARPC2 antibody [EPR8533] (AB133315)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling ARPC2 with purified ab133315 at 1 : 100 dilution (5.0 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARPC2 antibody [EPR8533] (AB133315)

Immunohistochemical analysis of paraffin embedded human kidney tissue labelled with ab133315 (unpurified) at 1/250 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ARPC2 antibody [EPR8533] (AB133315)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling ARPC2 with purified ab133315 at 1/50 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• WB

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Western blot - Anti-ARPC2 antibody [EPR8533] (AB133315)

All lanes:

Western blot - Anti-ARPC2 antibody [EPR8533] (ab133315) at 1/1000 dilution

Lane 1:

HeLa cell lysates at 10 µg

Lane 2:

293T cell lysates at 10 µg

Lane 3:

HT-1080 cell lysates at 10 µg

Lane 4:

Jurkat cell lysates at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 34 kDa

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• WB

Lab

Western blot - Anti-ARPC2 antibody [EPR8533] (AB133315)

All lanes:

Western blot - Anti-ARPC2 antibody [EPR8533] (ab133315) at 1/1000 dilution

Lane 1:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates at 20 µg

Lane 2:

Mouse heart lysates at 20 µg

Lane 3:

Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates at 20 µg

Lane 4:

Rat heart lysates 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: 34 kDa

Observed band size: 34 kDa

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• IP

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Immunoprecipitation - Anti-ARPC2 antibody [EPR8533] (AB133315)

ab133315 (purified ) at 1/30 dilution (2 µg) immunoprecipitating ARPC2 in HEK-293 whole cell lysate.
Lane 1 (input) : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10 µg
Lane 2 (+) : ab133315 & HEK-293 whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab133315 in HEK-293 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-ARPC2 antibody [EPR8533] (ab133315)

Predicted band size: 34 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-ARPC2 antibody [EPR8533] (AB133315)

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.