Anti-Rab3D antibody [EPR8106]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab3D antibody [EPR8106] (AB128997)

ab128997 at a 1/250 dilution staining Human Rab3D in stomach using Immunohistochemistry Formalin/PFA-fixed paraffin-emebedded tissue

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab3D antibody [EPR8106] (AB128997)

ab128997 at a 1/250 dilution staining Human Rab3D in clear cell carcinoma using Immunohistochemistry Formalin/PFA-fixed paraffin-emebedded tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Rab3D antibody [EPR8106] (AB128997)

Fluorescence multiplex immunohistochemical analysis of the human stomach (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-Chromogranin A (ab254557, gray; Opal™690), anti-MUC-6 (ab223846, green; Opal™520) and anti-Rab3D (ab128997, red; Opal™570) on human stomach. Panel B : anti-MUC-6 stained on mucous neck cells. Panel C : anti-Chromogranin A stained on neuroendocrine cells. Panel D : anti-Rab3D stained on Chief cells. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining : in the order of ab254557 (1/5000), ab223846 (1/1000), and ab128997 (1/10000) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Rab3D antibody [EPR8106] (AB128997)

Fluorescence multiplex immunohistochemical analysis of the human stomach (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-Mucin 5AC (ab3649 gray; Opal™690) anti-MUC-6 (ab223846 green; Opal™520) and anti-Rab3D (ab128997 red; Opal™570) on human stomach. Panel B : anti-MUC-6 stained on mucous neck cells. Panel C : anti-Mucin 5AC stained on surface mucous cells. Panel D : anti-Rab3D stained on Chief cells. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining : in the order of ab3649 (1/5000 dilution) ab223846 (1/1000 dilution) and ab128997 (1/10000 dilution) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0 epitope retrieval solution2) was used for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

• WB

Unknown

Western blot - Anti-Rab3D antibody [EPR8106] (AB128997)

All lanes:

Western blot - Anti-Rab3D antibody [EPR8106] (ab128997) at 1/10000 dilution

Lane 1:

BxPC-3 cell lysate at 10 µg

Lane 2:

T.T cell lysate at 10 µg

Lane 3:

A549 cell lysate at 10 µg

Lane 4:

HT-29 cell lysate at 10 µg

Secondary

All lanes:

Standard HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 24 kDa

Observed band size: 25 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Rab3D antibody [EPR8106] (AB128997)

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.