Anti-Lamin B1抗体-核Envelope Marker
参阅全部 Lamin B1 一抗
兔多克隆抗体to Lamin B1 -核Envelope Marker
Rabbit
From July 2024, QC testing of replenishment batches of this polyclonal changed. All tested and expected application and reactive species combinations are still covered by our Abcam product promise. However, we no longer test all applications. For more information on a specific batch, please contact our Scientific Support who will be happy to help.
适用于: ICC/IF, WB, IHC-Pmore details
与反应: Mouse, Rat, Human
预测可用于: Chicken, Pig, Xenopus laevis, Indian muntjac, Zebrafish
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
ICC/IF: HepG2 cells WB: HeLa, PC12 and NIH/3T3 whole cell lysate. Wild type HAP1 whole cell lysate. Wild type HAP1 nuclear lysate. Human Pancreatic cell line whole cell lysate. IHC-P: Human liver tissue. Human infantile fibromatosis tissue.
Lamin B1 and Lamin B antibodies are extremely useful as nuclear loading controls for use with nuclear extracts. When using Lamin B1 antibodies as nuclear loading controls, be aware that in apoptotic cells Lamin B1 is cleaved (Kottke TJ et al.). Lamin B1 will also be removed from a nuclear prep if the nuclear membranes are spun out. This antibody was designed to be a nuclear loading control however it has not yet been tested in appropriate lysates.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
浓度
100 µg 浓度为 1 mg/ml
Immunogen affinity purified
Lamin B1 and Lamin B antibodies are extremely useful as nuclear loading controls for use with nuclear extracts. When using Lamin B1 antibodies as nuclear loading controls, be aware that in apoptotic cells Lamin B1 is cleaved (Kottke TJ et al.). Lamin B1 will also be removed from a nuclear prep if the nuclear membranes are spun out. This antibody was designed to be a nuclear loading control however it has not yet been tested in appropriate lysates.
多克隆
IgG
Abpromise™承诺保证使用ab16048于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| ICC/IF | (27) | Use a concentration of 0.1 µg/ml. |
| WB | (66) | Use a concentration of 0.1 µg/ml. Detects a band of approximately 68 kDa (predicted molecular weight: 66 kDa). We recommend Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773). |
| IHC-P | (5) | Use a concentration of 0.1 - 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
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SwissProt: P14731 Chicken
SwissProt: P20700 Human
SwissProt: P14733 Mouse
Unigene: 89497 Human
Unigene: 4105 Mouse
Unigene: 11362 Rat
Entrez Gene: 396223 Chicken
Entrez Gene: 4001 Human
Entrez Gene: 16906 Mouse
Entrez Gene: 379745 Xenopus laevis
Entrez Gene: 195816 Zebrafish
Omim: 150340 Human
LMN antibody
LMN2 antibody
LMNB antibody
Lmnb1 antibody
LMNB1_HUMAN antibody
MGC111419 antibody
OTTHUMP00000159218 antibody
ADLD antibody
lamin B1 antibody
Lamin-B1 antibody
Immunocytochemistry - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)This image is courtesy of an anonymous Abreview
Immunocytochemistry analysis of formaldehyde-fixed NP40-permeabilized Mouse vascular smooth muscle cells staining with ab16048 at 1/100. Secondary antibody was Alexa Fluor® 488 donkey Anti-rabbit at 1/500 dilution. Samples were incubated with the primary antibody with 3% BSA in PBS for 12 hours at 4°C. Blocking was done using 3% BSA for 1 hour at 21°C.
Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
All lanes : Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048) at 1 µg/ml
Lane 1 : HeLa whole cell lysate
Lane 2 : PC12 whole cell lysate
Lane 3 : NIH 3T3 whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Predicted band size: 66 kDa
Blocking buffer: 3% Milk
Gel type: MOPS
Exposure Time: 2 minutes
Observed band size: 73 kDa
Additional bands: 46 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
Immunohistochemical analysis of formalin-fixed paraffin-embedded human liver labelling lamin B1 with ab16048 at a concentration of 0.2µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). ab16048 anti lamin B1 antibody was incubated at 37°C for 16min. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: empty lane
Lane 3: KO HAP1 LMNB1 whole cell lysate (20 µg)
Lane 4: empty lane
Lanes 1 - 4: Merged signal (red and green). Green - ab16048 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab16048 was shown to specifically react with LMNB1 (Lamin B1) in wild type HAP1 cells. No band was observed when LMNB1 (Lamin B1) knockout samples were used. Ab16048 LMNB1 (Lamin B1) and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 0.1 μg per mL and 1/10000 dilution respectively. Blots were developed with Goat 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 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
ab16048 staining Lamin B1 in HepG2 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab16048 at 0.1 µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 100% methanol (5 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
IHC image of Lamin B1 staining in Human normal Liver formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16048, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
All lanes : Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048) at 1 µg/ml
Lane 1 : Wild-type HAP1 nuclear lysate
Lane 2 : Lamin B1 knockout HAP1 nuclear lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 66 kDa
Observed band size: 68 kDawhy is the actual band size different from the predicted?
Lane 1: Wild-type HAP1 nuclear lysate (10 µg)
Lane 2: Lamin B1 knockout HAP1 nuclear lysate (10 µg)
Lanes 1 and 2: Green signal from target - ab16048 observed at 68 kDa. Red signal from loading control - ab10799 observed at 18 kDa.
ab16048 was shown to specifically react with lamin B1 in wild-type HAP1 cells. No band was observed knockout samples were used. Wild-type and lamin B1 knockout samples were subjected to SDS-PAGE. ab16048 and ab10799 (loading control to histone H3 at 0.1µg/mL) were both incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging.
Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
All lanes : Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048) at 1/1000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate with Mouse Lamin B1 peptide (ab16375) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Alexa fluor goat polyclonal to Rabbit IgG at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 66 kDa
Observed band size: 68-70 kDawhy is the actual band size different from the predicted?
Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)This image is courtesy of an anonymous Abreview
Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048) at 1/1000 dilution + Human Pancreatic cell line - whole cell lysate at 20 µg
Secondary
HRP conjugated goat anti-rabbit antibody at 1/2000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 66 kDa
Observed band size: 68 kDawhy is the actual band size different from the predicted?
Exposure time: 30 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)This image is courtesy of an anonymous Abreview
ab16048 staining Lamin B1 in human infantile fibromatosis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% FBS/BSA for 3 hours at room temperature; antigen retrieval was by heat mediation in Tris pH9. Samples were incubated with primary antibody (1/100 in TBS + 1% BSA + 1% FBS) for 16 hours. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)Image courtesy of Marilena Ciciarello and Patrizia Lavia, University 'La Sapienza' CNR, Italy
Human and mouse cells stained with ab16048 (1/500). The cells were fixed and permeabilized in 4% formaldehyde, 0.2% Tritonm X100 for 10 minutes at room temperature, then washed 3x in PBS.
A: HeLa cells + ab16048 (green)
B: HeLa cells counterstained with DAPI (blue)
C: 3T3 cells + ab16048 (green)
D: 3T3 cells counterstained with DAPI (blue)
Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)Image courtesy of Marilena Ciciarello and Patrizia Lavia, University 'La Sapienza' CNR, Italy
Human and mouse cells stained with ab16048 (1/500). The cells were fixed in 100% methanol for 6 minutes at -20°C, then washed once in PBS.
A: HeLa cells + ab16048 (green)
B: HeLa cells counterstained with DAPI (blue)
C: 3T3 cells + ab16048 (green)
D: 3T3 cells counterstained with DAPI (blue)
