Anti-HLA DMB antibody [EPR7981]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-HLA DMB antibody [EPR7981] (AB133640)

Intracellular Flow Cytometry analysis of permeabilized Raji cells labelling HLA DMB with ab133640 at 1/100 dilution (red). Rabbit IgG (negative control) shown in green.

• WB

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Western blot - Anti-HLA DMB antibody [EPR7981] (AB133640)

All lanes:

Western blot - Anti-HLA DMB antibody [EPR7981] (ab133640) at 1/1000 dilution

Lane 1:

Raji lysate at 10 µg

Lane 2:

Daudi lysate at 10 µg

Lane 3:

Human spleen lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 29 kDa

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• WB

Lab

Western blot - Anti-HLA DMB antibody [EPR7981] (AB133640)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. ab181602 was used as a loading control.

All lanes:

Western blot - Anti-HLA DMB antibody [EPR7981] (ab133640) at 1/10000 dilution

Lane 1:

293T (Human embryonic kidney epithelial cell) whole cell lysate with blank vector transfected at 15 µg

Lane 2:

293T (Human embryonic kidney epithelial cell) whole cell lysate with His tagged HLA DMB expression vector transfected at 15 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: 29 kDa

Observed band size: 29 kDa

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Exposure time: 1s

• OI-RD Scanning

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OI-RD Scanning - Anti-HLA DMB antibody [EPR7981] (AB133640)

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.

• WB

CiteAb

Western blot - Anti-HLA DMB antibody [EPR7981] (AB133640)

HLA DMB western blot using anti-HLA DMB antibody [EPR7981] ab133640. Publication image and figure legend from Liu, B., Feng, S., et al., 2019, BMC Nephrol, PubMed 31113397.

ab133640 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab133640 please see the product overview.

Upregulation of HLA-DMB expression on HPMCs after exposure to PYS or to HPG at day 3 and day 7. The mean of fluorescence intensity (MFI) of HLA-DMB staining on HPMCs after repeating exposure to PYS or to HPG at day 3 and day 7 compared to untreated control at day 0 (as a baseline) was measured by using flow cytometric analysis. The ratio of MFI on treated cells to its baseline was presented as fold change of HLA-DMB expression. a Data were presented as a typical histogram of HLA-DMB stain (solid line) in each group compared to the baseline (MFI : 126, dotted line), showing the fold change of HLA-DMB expression. b Data were presented as mean ± SD of four separate experiments (n = 4). p = 0.0036 (PYS vs. HPG, two-way ANOVA). *p = 0.0011 (PYS : day 3 vs. day 7, t-test). **p = 0.0032 (HPG : day 3 vs. day 7, t-test). (C) The total cellular levels of HLA-DMB staining on HPMCs after repeating exposure to PYS or to HPG at day 3 and day 7 compared to untreated control at day 0 were determined by Western blot analysis. Equal amount of protein (100 to 150 μg) extracted from whole cell pellets was fractioned by 10% of SDS-PAGE, and HLA-DMB protein bands were identified based on specifically binding of anti-HLA-DMB antibody, and their molecular size (26–28 kDa) (upper panel). The protein content in each sample was confirmed by re-probing the blot with anti-GAPDH antibody (middle panel) and was measured by densitometry. Imaging data are a representative of three separate experiments. The ratio of HLA-DMB band to GAPDH band from the same sample on the same blot was presented (bottom panel)

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