Anti-RAPGEF5 antibody [EPR6882]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RAPGEF5 antibody [EPR6882] (AB129008)

Immunocytochemistry/Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RAPGEF5 with Purified ab129008 at 1 : 500 dilution (0.24 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.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RAPGEF5 antibody [EPR6882] (AB129008)

ab129008, unpurified, at 1/100 dilution, staining RAPGEF5 in HEK-293 cells by Immunofluorescence

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-RAPGEF5 antibody [EPR6882] (AB129008)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells, labeling RAPGEF5 with Purified ab129008 at 1/20 dilution (10μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 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).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAPGEF5 antibody [EPR6882] (AB129008)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebrum tissue sections labeling RAPGEF5 with purified ab129008 at 1/100 dilution (1.19 μg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAPGEF5 antibody [EPR6882] (AB129008)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling RAPGEF5 with purified ab129008 at 1/100 dilution (1.19 µg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• WB

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Western blot - Anti-RAPGEF5 antibody [EPR6882] (AB129008)

All lanes:

Western blot - Anti-RAPGEF5 antibody [EPR6882] (ab129008) at 1/1000 dilution

Lane 1:

293T cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

K562 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled Goat anti-Rabbit IgG at 1/2000 dilution

Predicted band size: 68 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAPGEF5 antibody [EPR6882] (AB129008)

ab129008, unpurified, at 1/250 dilution, staining RAPGEF5 in paraffin embedded Human brain tissue by Immunohistochemistry.

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAPGEF5 antibody [EPR6882] (AB129008)

ab129008, unpurified, at 1/250 dilution, staining RAPGEF5 in paraffin embedded Human testis tissue by Immunohistochemistry.

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAPGEF5 antibody [EPR6882] (AB129008)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue sections labeling RAPGEF5 with purified ab129008 at 1/100 dilution (1.19 μg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• WB

Unknown

Western blot - Anti-RAPGEF5 antibody [EPR6882] (AB129008)

Blocking/Diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-RAPGEF5 antibody [EPR6882] (ab129008) at 1/1000 dilution

Lane 1:

293T (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 3:

HeLa (Human cervix adenocarcinoma epithelial cel) whole cell lysate at 20 µg

Lane 4:

Mouse brain lysate at 20 µg

Lane 5:

Rat brain lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 68 kDa

Observed band size: 68 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-RAPGEF5 antibody [EPR6882] (AB129008)

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.