Anti-67kDa Laminin Receptor antibody [EPR8469]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-67kDa Laminin Receptor antibody [EPR8469] (AB133645)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labellig 67kDa Laminin Receptor with ab133645 at 1/100 dilution.

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-67kDa Laminin Receptor antibody [EPR8469] (AB133645)

ab133645 staining 67kDa Laminin Receptor in human gastric carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-67kDa Laminin Receptor antibody [EPR8469] (AB133645)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labelling 67kDa Laminin Receptor with ab133645 at 1/100 dilution.

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

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-67kDa Laminin Receptor antibody [EPR8469] (AB133645)

ab133645 staining 67kDa Laminin Receptor in the human cell line MCF7 (Human breast adenocarcinoma cell line) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control : Rabbit monoclonal IgG (Black)

Unlabelled control : Cell without incubation with primary antibody and secondary antibody (Blue)

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-67kDa Laminin Receptor antibody [EPR8469] (AB133645)

ab133645 staining 67kDa Laminin Receptor in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. ab195889 was used as a counterstain for primary antibody ab133645 at 1/2000. DAPI was used as a nuclear counterstain and PBS as a negative control.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-67kDa Laminin Receptor antibody [EPR8469] (AB133645)

Overlay histogram showing MCF7 cells stained with ab133645 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab133645, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• IP

Lab

Immunoprecipitation - Anti-67kDa Laminin Receptor antibody [EPR8469] (AB133645)

ab133645 immunoprecipitating 67kDa Laminin Receptor. 10μg of cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/1000.

Lane 1 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate (10ug)
Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab133645 in K562(Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate

All lanes:

Immunoprecipitation - Anti-67kDa Laminin Receptor antibody [EPR8469] (ab133645)

Predicted band size: 33 kDa

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• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-67kDa Laminin Receptor antibody [EPR8469] (AB133645)

ab133645 staining Prohibitin in mouse kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-67kDa Laminin Receptor antibody [EPR8469] (AB133645)

ab133645 staining 67kDa Laminin Receptor in mouse kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-67kDa Laminin Receptor antibody [EPR8469] (AB133645)

ab133645 staining 67kDa Laminin Receptor in rat kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

• WB

Lab

Western blot - Anti-67kDa Laminin Receptor antibody [EPR8469] (AB133645)

Diluting and blocking buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-67kDa Laminin Receptor antibody [EPR8469] (ab133645) at 1/2000 dilution

Lane 1:

K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

C6 (Rat glial tumor cell line) whole cell lysate 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: 33 kDa

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• WB

Lab

Western blot - Anti-67kDa Laminin Receptor antibody [EPR8469] (AB133645)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-67kDa Laminin Receptor antibody [EPR8469] (ab133645) at 1/10000 dilution

Lane 1:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg

Lane 2:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate 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: 33 kDa,54 kDa

Observed band size: 54 kDa

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• WB

AbReview42020****

Western blot - Anti-67kDa Laminin Receptor antibody [EPR8469] (AB133645)

All lanes:

Western blot - Anti-67kDa Laminin Receptor antibody [EPR8469] (ab133645) at 1/1000 dilution

All lanes:

Mouse intestinal mucosa whole tissue lysate at 30 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG polyclonal at 1/2500 dilution

Predicted band size: 33 kDa

Observed band size: 55 kDa

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Exposure time: 1min

This image is courtesy of an anonymous Abreview

• WB

Unknown

Western blot - Anti-67kDa Laminin Receptor antibody [EPR8469] (AB133645)

All lanes:

Western blot - Anti-67kDa Laminin Receptor antibody [EPR8469] (ab133645) at 1/1000 dilution

Lane 1:

K562 cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

HepG2 cell lysate at 10 µg

Lane 4:

C6 cell lysate at 10 µg

Lane 5:

RAW 264.7 cell lysate at 10 µg

Lane 6:

PC-12 cell lysate at 10 µg

Lane 7:

NIH/3T3 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

false

• WB

CiteAb

Western blot - Anti-67kDa Laminin Receptor antibody [EPR8469] (AB133645)

67kDa Laminin Receptor western blot using anti-67kDa Laminin Receptor antibody [EPR8469] ab133645. Publication image and figure legend from Gao, S., Li, C., et al., 2017, Sci Rep, PubMed 28211523.

ab133645 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 ab133645 please see the product overview.

PEDF regulated hypoxia-induced macrophages polorization via ATGL.(a,b) Immunofluorescence analysis of ATGL and LR expression on macrophages of retinas in the OIR model by ATGL plus F4/80 (a) or LR plus F4/80 (b) staining. Scale bars represent 50 μm. (c) Immunofluorescence analysis of ATGL and LR expression in macrophage RAW264.7 and BMDMs with or without hypoxia treatment. Scale bars represent 10 μm. (d) Western blot analysis of ATGL and LR expression in macrophages RAW264.7 and BMDMs with or without hypoxia treatment. (e,f) Quantification of iNOS and Arg-1 expression in BMDMs induced by hypoxia prior to stimulation with PEDF and/or ATGL specific inhibitor BEL/ATGL selective inhibitor MAFP/LR antibody MluC5. ***p < 0.001, P values were analyzed by one-way ANOVA. All data are representative of three independent experiments and are means ± SEM.

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