WB
Western blot analysis of LAMC2 on different lysates with Rabbit anti-LAMC2 antibody at 1/5,000 dilution. Lane 1: A431 cell lysate (20 µg/Lane), Lane 2: A549 cell lysate (low expression) (20 µg/Lane), Lane 3: HepG2 cell lysate (low expression) (20 µg/Lane), Lane 4: HaCaT cell lysate (low expression) (20 µg/Lane), Lane 5: NCI-H441 cell lysate (20 µg/Lane), Lane 6: Mouse skin tissue lysate (40 µg/Lane), Lane 7: Mouse stomach tissue lysate (40 µg/Lane), Lane 8: Rat skin tissue lysate (40 µg/Lane), Lane 9: Rat stomach tissue lysate (40 µg/Lane), Exposure time: 2 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-LAMC2 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-LAMC2 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-LAMC2 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of A431 (positive) and A549 (low expression) labeling LAMC2 with Rabbit anti-LAMC2 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-LAMC2 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of A549 (left, low expression) and A431 (right, positive) cells labeling LAMC2. Cells were fixed and permeabilized. Then stained with the primary antibody (1/5,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
LAMC2 was immunoprecipitated from 0.2 mg A431 cell lysate with Rabbit anti-LAMC2 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-LAMC2 antibody at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: A431 cell lysate (input), Lane 2: Rabbit anti-LAMC2 antibody IP in A431 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-LAMC2 antibody in A431 cell lysate, Blocking/Dilution buffer: primary antibody dilution, Exposure time: 2 seconds.| Product Name | LAMC2 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human LAMC2 aa 550-1,193. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:5000 IHC:1:200 ICC/IF:1:100 FC:1:5000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | LAMC2 |
|---|---|
| Gene Synonyms | B2T CSF EBR2 BM600 EBR2A JEB3A JEB3B LAMB2T LAMNB2 |
| Gene Full Name | laminin subunit gamma 2 |
| Gene Summary | Laminins, a family of extracellular matrix glycoproteins, are the major noncollagenous constituent of basement membranes. They have been implicated in a wide variety of biological processes including cell adhesion, differentiation, migration, signaling, neurite outgrowth and metastasis. Laminins, composed of 3 non identical chains: laminin alpha, beta and gamma (formerly A, B1, and B2, respectively), have a cruciform structure consisting of 3 short arms, each formed by a different chain, and a long arm composed of all 3 chains. Each laminin chain is a multidomain protein encoded by a distinct gene. Several isoforms of each chain have been described. Different alpha, beta and gamma chain isomers combine to give rise to different heterotrimeric laminin isoforms which are designated by Arabic numerals in the order of their discovery, i.e. alpha1beta1gamma1 heterotrimer is laminin 1. The biological functions of the different chains and trimer molecules are largely unknown, but some of the chains have been shown to differ with respect to their tissue distribution, presumably reflecting diverse functions in vivo. This gene encodes the gamma chain isoform laminin, gamma 2. The gamma 2 chain, formerly thought to be a truncated version of beta chain (B2t), is highly homologous to the gamma 1 chain; however, it lacks domain VI, and domains V, IV and III are shorter. It is expressed in several fetal tissues but differently from gamma 1, and is specifically localized to epithelial cells in skin, lung and kidney. The gamma 2 chain together with alpha 3 and beta 3 chains constitute laminin 5 (earlier known as kalinin), which is an integral part of the anchoring filaments that connect epithelial cells to the underlying basement membrane. The epithelium-specific expression of the gamma 2 chain implied its role as an epithelium attachment molecule, and mutations in this gene have been associated with junctional epidermolysis bullosa, a skin disease characterized by blisters due to disruption of the epidermal-dermal junction. Two transcript variants resulting from alternative splicing of the 3' terminal exon, and encoding different isoforms of gamma 2 chain, have been described. The two variants are differentially expressed in embryonic tissues, however, the biological significance of the two forms is not known. Transcript variants utilizing alternative polyA_signal have also been noted in literature. [provided by RefSeq, Aug 2011] |
| Alternative Names | 3918 antibody B2T antibody BM600 antibody Cell-scattering factor 140 kDa subunit antibody CSF 140 kDa subunit antibody CSF antibody EBR2 antibody EBR2A antibody Epiligrin subunit gamma antibody Kalinin subunit gamma antibody 3918 antibody B2T antibody BM600 antibody Cell-scattering factor 140 kDa subunit antibody CSF 140 kDa subunit antibody CSF antibody EBR2 antibody EBR2A antibody Epiligrin subunit gamma antibody Kalinin subunit gamma antibody Kalinin/nicein/epiligrin 100 kDa subunit antibody Ladsin 140 kDa subunit antibody LAMB2T antibody LAMC2 antibody LAMC2_HUMAN antibody Laminin 5 gamma 2 subunit antibody Laminin B2t chain antibody Laminin gamma 2 antibody laminin gamma 2 chain antibody Laminin subunit gamma-2 antibody Laminin-5 subunit gamma antibody LAMNB2 antibody Large adhesive scatter factor 140 kDa subunit antibody MGC138491 antibody MGC141938 antibody Nicein subunit gamma antibody NICEIN-100KDA antibody |
| Molecular Weight(MW) | 131kDa(Observed band size:100/140kDa) |
| Cellular Localization | Secreted, extracellular space, extracellular matrix, basement membrane. |
WB
Western blot analysis of LAMC2 on different lysates with Rabbit anti-LAMC2 antibody at 1/5,000 dilution. Lane 1: A431 cell lysate (20 µg/Lane), Lane 2: A549 cell lysate (low expression) (20 µg/Lane), Lane 3: HepG2 cell lysate (low expression) (20 µg/Lane), Lane 4: HaCaT cell lysate (low expression) (20 µg/Lane), Lane 5: NCI-H441 cell lysate (20 µg/Lane), Lane 6: Mouse skin tissue lysate (40 µg/Lane), Lane 7: Mouse stomach tissue lysate (40 µg/Lane), Lane 8: Rat skin tissue lysate (40 µg/Lane), Lane 9: Rat stomach tissue lysate (40 µg/Lane), Exposure time: 2 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-LAMC2 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-LAMC2 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-LAMC2 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of A431 (positive) and A549 (low expression) labeling LAMC2 with Rabbit anti-LAMC2 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-LAMC2 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of A549 (left, low expression) and A431 (right, positive) cells labeling LAMC2. Cells were fixed and permeabilized. Then stained with the primary antibody (1/5,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
LAMC2 was immunoprecipitated from 0.2 mg A431 cell lysate with Rabbit anti-LAMC2 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-LAMC2 antibody at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: A431 cell lysate (input), Lane 2: Rabbit anti-LAMC2 antibody IP in A431 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-LAMC2 antibody in A431 cell lysate, Blocking/Dilution buffer: primary antibody dilution, Exposure time: 2 seconds.| Application Notes | WB:1:5000 IHC:1:200 ICC/IF:1:100 FC:1:5000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
