Anti-RAB7 antibody [EPR7588(B)] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB7 antibody [EPR7588(B)] - BSA and Azide free (AB238990)

Immunohistochemical staining of paraffin-embedded human bladder carcinoma sections labelling RAB7 with purified ab126712 at dilution of 1/70. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counter-stained 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126712).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB7 antibody [EPR7588(B)] - BSA and Azide free (AB238990)

Immunohistochemical analysis of paraffin-embedded Human colon adenocarcinoma tissue labelling Rab7A with unpurified ab126712 at dilution of 1/50.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126712).

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

• WB

Lab

Western blot - Anti-RAB7 antibody [EPR7588(B)] - BSA and Azide free (AB238990)

Lanes 1 - 2 : Merged signal (red and green). Green - ab126712 observed at 23 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab126712 was shown to react with RAB7 in wild-type HeLa cells in Western blot with loss of signal observed in RAB7A knockout cell line ab255423 (RAB7A knockout cell lysate ab263831). Wild-type HeLa and RAB7A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab126712 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-RAB7 antibody [EPR7588(B)] - BSA and Azide free (ab238990) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

RAB7A knockout HeLa cell lysate at 20 µg

Observed band size: 23 kDa

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• WB

Lab

Western blot - Anti-RAB7 antibody [EPR7588(B)] - BSA and Azide free (AB238990)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : RAB7 knockout HAP1 cell lysate (20 μg)
Lane 3 : A431 cell lysate (20 μg)
Lane 4 : Human fetal brain tissue lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab126712 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab126712 was shown to recognize RAB7 when RAB7 knockout samples were used, along with additional cross-reactive bands. Wild-type and RAB7 knockout samples were subjected to SDS-PAGE. ab126712 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 and incubated overnight at 4°C. Blots were developed withGoat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab126712).

All lanes:

Western blot - Anti-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (<a href='/en-us/products/primary-antibodies/rab7-antibody-epr7588b-late-endosome-marker-ab126712'>ab126712</a>)

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• OI-RD Scanning

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OI-RD Scanning - Anti-RAB7 antibody [EPR7588(B)] - BSA and Azide free (AB238990)

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.