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human lambda light chain kit  (Fortis Life Sciences)


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    Fortis Life Sciences human lambda light chain kit
    A . RPMI-8226, ALMC-2, MM.1S, and U266 cells were incubated for 48 hours in the presence or absence of lovastatin ( Lov , 2-50 μM) or DGBP (2-50 μM). Immunoblot analysis of p62, Atg3 and β-tubulin (loading control) is shown. The relative intensity ( Rel Int) values underneath the p62 and Atg3 blots represent relative levels as determined by densitometric analysis of Atg3 or p62 (relative to tubulin) of treated cells compared to control cells. The gels are representative of 3-4 independent experiments. B . ALMC-2 cells were incubated for 48 hours in the presence of 10 μM lovastatin ( Lov ) and/or mevalonate ( Mev , 5 mM), FPP (10 μM), or GGPP (10 μM) for 48 hrs. Densitometric analysis of p62 or Atg3 levels (normalized to β-tubulin levels) for the combination treatments normalized to untreated cells (control) is shown. Data are displayed as average +/− standard deviation (n=3 independent experiments). * denotes p-value <0.05 from two-sided t-testing. C . RPMI-8226 and ALMC-2 cells were incubated for 48 hours in the presence or absence of 10 μM lovastatin ( Lov ), 10 μM DGBP, 10 μM GGTI-2133 ( GGTI ), 10 mM 3-PEHPC ( 3P ), or the combination of 10 μM GGTI-2133 and 10 mM 3-PEHPC ( GGTI + 3P ). Immunoblot analysis of p62, Atg3 and β-tubulin (loading control) is shown. The gels are representative of 3-4 independent experiments. D . RPMI-8226 and ALMC-2 cells were incubated for 48 hours in the presence or absence of 10 μM lovastatin ( Lov ), 10 μM DGBP, 10 mM 3-PEHPC ( 3P ), or 5 nM bafilomycin A1 ( Baf ) with or without protease inhibitors (10 μg/mL E64d and pepstatin A, prot ). Immunoblot analysis of LC3 and β-tubulin (loading control) is shown. The gels are representative of 3 independent experiments. E <t>.</t> <t>Intracellular</t> <t>lambda</t> light chain levels were measured via ELISA. Data are expressed as a percentage of control (mean +/− standard deviation of 3 independent experiments). Two-sided t-testing was performed to compare the IBP inhibitor alone vs. in combination with the protease inhibitors. No significant differences between the treatment groups were observed.
    Human Lambda Light Chain Kit, supplied by Fortis Life Sciences, used in various techniques. Bioz Stars score: 93/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/free+human+ig+lambda+light+chains/Human+Lambda+Free+Light+Chain/pmc04747172-164-1-7
    Average 93 stars, based on 36 article reviews
    human lambda light chain kit - by Bioz Stars, 2026-11
    93/100 stars

    Images

    1) Product Images from "Mechanisms for autophagy modulation by isoprenoid biosynthetic pathway inhibitors in multiple myeloma cells"

    Article Title: Mechanisms for autophagy modulation by isoprenoid biosynthetic pathway inhibitors in multiple myeloma cells

    Journal: Oncotarget

    doi:

    A . RPMI-8226, ALMC-2, MM.1S, and U266 cells were incubated for 48 hours in the presence or absence of lovastatin ( Lov , 2-50 μM) or DGBP (2-50 μM). Immunoblot analysis of p62, Atg3 and β-tubulin (loading control) is shown. The relative intensity ( Rel Int) values underneath the p62 and Atg3 blots represent relative levels as determined by densitometric analysis of Atg3 or p62 (relative to tubulin) of treated cells compared to control cells. The gels are representative of 3-4 independent experiments. B . ALMC-2 cells were incubated for 48 hours in the presence of 10 μM lovastatin ( Lov ) and/or mevalonate ( Mev , 5 mM), FPP (10 μM), or GGPP (10 μM) for 48 hrs. Densitometric analysis of p62 or Atg3 levels (normalized to β-tubulin levels) for the combination treatments normalized to untreated cells (control) is shown. Data are displayed as average +/− standard deviation (n=3 independent experiments). * denotes p-value <0.05 from two-sided t-testing. C . RPMI-8226 and ALMC-2 cells were incubated for 48 hours in the presence or absence of 10 μM lovastatin ( Lov ), 10 μM DGBP, 10 μM GGTI-2133 ( GGTI ), 10 mM 3-PEHPC ( 3P ), or the combination of 10 μM GGTI-2133 and 10 mM 3-PEHPC ( GGTI + 3P ). Immunoblot analysis of p62, Atg3 and β-tubulin (loading control) is shown. The gels are representative of 3-4 independent experiments. D . RPMI-8226 and ALMC-2 cells were incubated for 48 hours in the presence or absence of 10 μM lovastatin ( Lov ), 10 μM DGBP, 10 mM 3-PEHPC ( 3P ), or 5 nM bafilomycin A1 ( Baf ) with or without protease inhibitors (10 μg/mL E64d and pepstatin A, prot ). Immunoblot analysis of LC3 and β-tubulin (loading control) is shown. The gels are representative of 3 independent experiments. E . Intracellular lambda light chain levels were measured via ELISA. Data are expressed as a percentage of control (mean +/− standard deviation of 3 independent experiments). Two-sided t-testing was performed to compare the IBP inhibitor alone vs. in combination with the protease inhibitors. No significant differences between the treatment groups were observed.
    Figure Legend Snippet: A . RPMI-8226, ALMC-2, MM.1S, and U266 cells were incubated for 48 hours in the presence or absence of lovastatin ( Lov , 2-50 μM) or DGBP (2-50 μM). Immunoblot analysis of p62, Atg3 and β-tubulin (loading control) is shown. The relative intensity ( Rel Int) values underneath the p62 and Atg3 blots represent relative levels as determined by densitometric analysis of Atg3 or p62 (relative to tubulin) of treated cells compared to control cells. The gels are representative of 3-4 independent experiments. B . ALMC-2 cells were incubated for 48 hours in the presence of 10 μM lovastatin ( Lov ) and/or mevalonate ( Mev , 5 mM), FPP (10 μM), or GGPP (10 μM) for 48 hrs. Densitometric analysis of p62 or Atg3 levels (normalized to β-tubulin levels) for the combination treatments normalized to untreated cells (control) is shown. Data are displayed as average +/− standard deviation (n=3 independent experiments). * denotes p-value <0.05 from two-sided t-testing. C . RPMI-8226 and ALMC-2 cells were incubated for 48 hours in the presence or absence of 10 μM lovastatin ( Lov ), 10 μM DGBP, 10 μM GGTI-2133 ( GGTI ), 10 mM 3-PEHPC ( 3P ), or the combination of 10 μM GGTI-2133 and 10 mM 3-PEHPC ( GGTI + 3P ). Immunoblot analysis of p62, Atg3 and β-tubulin (loading control) is shown. The gels are representative of 3-4 independent experiments. D . RPMI-8226 and ALMC-2 cells were incubated for 48 hours in the presence or absence of 10 μM lovastatin ( Lov ), 10 μM DGBP, 10 mM 3-PEHPC ( 3P ), or 5 nM bafilomycin A1 ( Baf ) with or without protease inhibitors (10 μg/mL E64d and pepstatin A, prot ). Immunoblot analysis of LC3 and β-tubulin (loading control) is shown. The gels are representative of 3 independent experiments. E . Intracellular lambda light chain levels were measured via ELISA. Data are expressed as a percentage of control (mean +/− standard deviation of 3 independent experiments). Two-sided t-testing was performed to compare the IBP inhibitor alone vs. in combination with the protease inhibitors. No significant differences between the treatment groups were observed.

    Techniques Used: Incubation, Western Blot, Standard Deviation, Enzyme-linked Immunosorbent Assay

    RPMI-8226 cells were incubated with 2 nM bafilomycinA1, 10 μM lovastatin, or 5 mM 3-PEHPC for 48 hours. Staining was performed as described in Materials and Methods, using antibodies directed against lambda light chain (red) and PDI as an ER marker (green) in A . and RCAS1 as a Golgi marker (green) in B . DAPI was used for nuclear staining (blue). Colocalization was determined using the RGB plot profile tool in ImageJ and indicated by similarity of the patterns of red and green peaks. Scale bar 10μm.
    Figure Legend Snippet: RPMI-8226 cells were incubated with 2 nM bafilomycinA1, 10 μM lovastatin, or 5 mM 3-PEHPC for 48 hours. Staining was performed as described in Materials and Methods, using antibodies directed against lambda light chain (red) and PDI as an ER marker (green) in A . and RCAS1 as a Golgi marker (green) in B . DAPI was used for nuclear staining (blue). Colocalization was determined using the RGB plot profile tool in ImageJ and indicated by similarity of the patterns of red and green peaks. Scale bar 10μm.

    Techniques Used: Incubation, Staining, Marker

    A . RPMI-8226 cells were incubated with 2 nM bafilomycin A1 ( Baf ), 10 μM lovastatin ( Lov ), or 5 mM 3-PEHPC ( 3P ) for 48 h or 10 nM bortezomib for 24 h. Staining was performed as described in Materials and Methods, using Proteostat (Enzo Life Sciences, Inc.) (green) to visualize aggresomes and an antibody directed against lambda light chain (red). DAPI was used for nuclear staining (blue). Colocalization was determined using the RGB plot profile tool in ImageJ. Scale bar 10 μm. B . The average number of aggresomes per cell was quantified using the GFP-LC3 macro for ImageJ (mean ± SEM). **** denotes P <0.0001 from unpaired two-tailed t- test comparing treated with control cells.
    Figure Legend Snippet: A . RPMI-8226 cells were incubated with 2 nM bafilomycin A1 ( Baf ), 10 μM lovastatin ( Lov ), or 5 mM 3-PEHPC ( 3P ) for 48 h or 10 nM bortezomib for 24 h. Staining was performed as described in Materials and Methods, using Proteostat (Enzo Life Sciences, Inc.) (green) to visualize aggresomes and an antibody directed against lambda light chain (red). DAPI was used for nuclear staining (blue). Colocalization was determined using the RGB plot profile tool in ImageJ. Scale bar 10 μm. B . The average number of aggresomes per cell was quantified using the GFP-LC3 macro for ImageJ (mean ± SEM). **** denotes P <0.0001 from unpaired two-tailed t- test comparing treated with control cells.

    Techniques Used: Incubation, Staining, Two Tailed Test

    U266 cells were treated with 2 nM bafilomycin A1 ( Baf ), 10 μM lovastatin ( Lov ) or 10 mM 3-PEHPC for 48 hours before processing for staining with LC3. A . The average number of LC3 puncta per cell and B . the average size of each punctate structure in pixels was quantified using the GFP-LC3 macro for ImageJ (mean ± SEM) (n>175). The **** denotes p <0.0001 from unpaired two-tailed t- test comparing treated with control cells. C . Representative confocal maximum projections are shown for each treatment with LC3 in green and DAPI staining the nucleus in blue. D . RPMI-8226 cells were incubated with 2 nM bafilomycin A1, 10μM lovastatin, 5mM 3-PEHPC or a co-treatment with bafilomycin A1 ( Baf ) and lovastatin ( Lov ) for 48 hours. Staining was performed as described in Materials and Methods, using CytoID (Enzo Life Sciences, Inc., Farmingdale, NY) to visualize autophagosomes and an antibody directed against lambda light chain. DAPI was used for nuclear staining. Colocalization was determined using the RGB plot profile tool in ImageJ. Scale bar 10μm.
    Figure Legend Snippet: U266 cells were treated with 2 nM bafilomycin A1 ( Baf ), 10 μM lovastatin ( Lov ) or 10 mM 3-PEHPC for 48 hours before processing for staining with LC3. A . The average number of LC3 puncta per cell and B . the average size of each punctate structure in pixels was quantified using the GFP-LC3 macro for ImageJ (mean ± SEM) (n>175). The **** denotes p <0.0001 from unpaired two-tailed t- test comparing treated with control cells. C . Representative confocal maximum projections are shown for each treatment with LC3 in green and DAPI staining the nucleus in blue. D . RPMI-8226 cells were incubated with 2 nM bafilomycin A1, 10μM lovastatin, 5mM 3-PEHPC or a co-treatment with bafilomycin A1 ( Baf ) and lovastatin ( Lov ) for 48 hours. Staining was performed as described in Materials and Methods, using CytoID (Enzo Life Sciences, Inc., Farmingdale, NY) to visualize autophagosomes and an antibody directed against lambda light chain. DAPI was used for nuclear staining. Colocalization was determined using the RGB plot profile tool in ImageJ. Scale bar 10μm.

    Techniques Used: Staining, Two Tailed Test, Incubation

    Related Articles

    Expressing:

    Article Title: Anti-lambda myeloma antigen (LMA) binding proteins to treat LMA-expressing cancer and autoimmune disorders
    Article Snippet: .. Similarly, human anti-LMA antibodies (6A1, 13H3 and 4A1) demonstrated high and selective affinity to human multiple myeloma cell lines expressing lambda-light chain isotypes and not to kappa-light chain expressing JJN3 cell line (FIG. 4) while clones 18E8, 7F11 and 18F9 demonstrated selective affinity to commercial lambda-light chain (Bethyl) (FIG. 2B). ..

    Clone Assay:

    Article Title: Anti-lambda myeloma antigen (LMA) binding proteins to treat LMA-expressing cancer and autoimmune disorders
    Article Snippet: .. Similarly, human anti-LMA antibodies (6A1, 13H3 and 4A1) demonstrated high and selective affinity to human multiple myeloma cell lines expressing lambda-light chain isotypes and not to kappa-light chain expressing JJN3 cell line (FIG. 4) while clones 18E8, 7F11 and 18F9 demonstrated selective affinity to commercial lambda-light chain (Bethyl) (FIG. 2B). ..

    Enzyme-linked Immunosorbent Assay:

    Article Title: Heterohybridomas producing human immunoglobulin light chains using CD138-selected bone marrow cells.
    Article Snippet: Background: Light chain research is hampered by lack of mammalian cell lines producing human light chains (FLC).. Therefore, we used heterohybridoma (HH) technology to produce clones making FLC thereby providing tools to study light chain behavior.. Methods: Marrow CD138+ cells from patients with multiple myeloma (MM) and polyclonal gammopathy (PG) were selected, fused with B5-6 T cells and cultured in hypoxanthine-aminopterin-thymidine medium (HAT).

    Article Title: Treatment of al amyloidosis with melflufen
    Article Snippet: .. Levels of kappa and lambda light chains as well as BCMA from the serum was analysed on days 9 and 16 post tumor inoculation using commercial ELISA kits (below) and manufacturer's protocols: Human Kappa ELISA (Bethyl Laboratories, #E80-115) Human Lambda ELISA (Bethyl Laboratories, #E80-116) Human BCMA ELISA (R&D Systems, #DY193). ..

    Article Title: Therapeutic implications of mitochondrial stress-induced proteasome inhibitor resistance in multiple myeloma.
    Article Snippet: .. Lysates of the cell pellets were prepared in RIPA buffer, and the lambda and kappa light chain antibodies were measured using Bethyl Human Lambda ELISA Quantitation Set (E80-116) and Bethyl Human Kappa ELISA Quantitation Set (#E80-115) following the manufacturer’s protocol. .. Bacterial clones containing ATF4 short hairpin RNAs were obtained from Sigma-Aldrich (TRCN0000013573 and TRCN0000013574) and plated on LB agar plates containing ampicillin.

    Article Title: Heterohybridomas producing human immunoglobulin light chains using CD138-selected bone marrow cells
    Article Snippet: .. Heterohybridoma supernatants for ELISA were evaluated in quantitative sandwich ELISA for human IgA, IgG, IgM, κ and λ light chains (Bethyl Laboratories, Montgomery, TX) according to the manufacturers’ instructions. ..

    Article Title: Therapeutic implications of mitochondrial stress–induced proteasome inhibitor resistance in multiple myeloma
    Article Snippet: .. Lysates of the cell pellets were prepared in RIPA buffer, and the lambda and kappa light chain antibodies were measured using Bethyl Human Lambda ELISA Quantitation Set (E80-116) and Bethyl Human Kappa ELISA Quantitation Set (#E80-115) following the manufacturer’s protocol. .. Lysates of the cell pellets were prepared in RIPA buffer, and the lambda and kappa light chain antibodies were measured using Bethyl Human Lambda ELISA Quantitation Set (E80-116) and Bethyl Human Kappa ELISA Quantitation Set (#E80-115) following the manufacturer’s protocol.

    Article Title: A non-ligand surrogate agonist antibody that enhances canonical Wnt signaling and bone regeneration
    Article Snippet: .. One week post treatment, mice were anesthetized and scanned by micro-CT. One week post CT scanning, blood and femurs were collected from the mice, and free human Ig-lambda light chains in serum was assessed using a Human Lambda ELISA Kit (Bethyl Laboratories) according to manufacturer’s instructions. .. One week post treatment, mice were anesthetized and scanned by micro-CT. One week post CT scanning, blood and femurs were collected from the mice, and free human Ig-lambda light chains in serum was assessed using a Human Lambda ELISA Kit (Bethyl Laboratories) according to manufacturer’s instructions.

    Sandwich ELISA:

    Article Title: Heterohybridomas producing human immunoglobulin light chains using CD138-selected bone marrow cells.
    Article Snippet: Background: Light chain research is hampered by lack of mammalian cell lines producing human light chains (FLC).. Therefore, we used heterohybridoma (HH) technology to produce clones making FLC thereby providing tools to study light chain behavior.. Methods: Marrow CD138+ cells from patients with multiple myeloma (MM) and polyclonal gammopathy (PG) were selected, fused with B5-6 T cells and cultured in hypoxanthine-aminopterin-thymidine medium (HAT).

    Article Title: Heterohybridomas producing human immunoglobulin light chains using CD138-selected bone marrow cells
    Article Snippet: .. Heterohybridoma supernatants for ELISA were evaluated in quantitative sandwich ELISA for human IgA, IgG, IgM, κ and λ light chains (Bethyl Laboratories, Montgomery, TX) according to the manufacturers’ instructions. ..

    Quantitation Assay:

    Article Title: Therapeutic implications of mitochondrial stress-induced proteasome inhibitor resistance in multiple myeloma.
    Article Snippet: .. Lysates of the cell pellets were prepared in RIPA buffer, and the lambda and kappa light chain antibodies were measured using Bethyl Human Lambda ELISA Quantitation Set (E80-116) and Bethyl Human Kappa ELISA Quantitation Set (#E80-115) following the manufacturer’s protocol. .. Bacterial clones containing ATF4 short hairpin RNAs were obtained from Sigma-Aldrich (TRCN0000013573 and TRCN0000013574) and plated on LB agar plates containing ampicillin.

    Article Title: Therapeutic implications of mitochondrial stress–induced proteasome inhibitor resistance in multiple myeloma
    Article Snippet: .. Lysates of the cell pellets were prepared in RIPA buffer, and the lambda and kappa light chain antibodies were measured using Bethyl Human Lambda ELISA Quantitation Set (E80-116) and Bethyl Human Kappa ELISA Quantitation Set (#E80-115) following the manufacturer’s protocol. .. Lysates of the cell pellets were prepared in RIPA buffer, and the lambda and kappa light chain antibodies were measured using Bethyl Human Lambda ELISA Quantitation Set (E80-116) and Bethyl Human Kappa ELISA Quantitation Set (#E80-115) following the manufacturer’s protocol.

    Micro-CT:

    Article Title: A non-ligand surrogate agonist antibody that enhances canonical Wnt signaling and bone regeneration
    Article Snippet: .. One week post treatment, mice were anesthetized and scanned by micro-CT. One week post CT scanning, blood and femurs were collected from the mice, and free human Ig-lambda light chains in serum was assessed using a Human Lambda ELISA Kit (Bethyl Laboratories) according to manufacturer’s instructions. .. One week post treatment, mice were anesthetized and scanned by micro-CT. One week post CT scanning, blood and femurs were collected from the mice, and free human Ig-lambda light chains in serum was assessed using a Human Lambda ELISA Kit (Bethyl Laboratories) according to manufacturer’s instructions.



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    Image Search Results


    Runx2 knockdown inhibits MM progression in vivo. (A) Expression of Runx2 in 2 Runx2 k/d 5TGM1 cell lines (Runx2 shRNA #90– and Runx2 shRNA #91–transfected 5TGM1 cells) was probed by western blot. Both Runx2 shRNA #90 and #91 decreased the expression of Runx2 compared with NT control. (B) Serum IgG2bκ was measured by ELISA 6 weeks after IV injection of NT control or Runx2 k/d 5TGM1 cells. Error bars represent mean ± SEM (n = 10 animals per group). Significant differences between groups are indicated by P value. (C) H&E-stained bone sections from mice injected IV with either NT control or Runx2 k/d 5TGM1 cells. Tumors were present in mice injected with NT cells, but not in the mice injected with Runx2 k/d cells (original magnification, ×100). Inset, Abundant myeloma cells in NT-bearing mice compared with Runx2 k/d 5TGM1 cell-injected mice. (D) Survival was significantly increased in mice injected IV with 5TGM1 Runx2 k/d cells (clone #90 and #91) compared with those injected with NT 5TGM1 cells. (E) Six weeks after intratibial injection of Runx2 k/d or NT 5TGM1 cells, levels of serum IgG2bκ were measured by ELISA. Error bars represent mean ± SEM (n = 10 animals per group). Significant differences between groups are indicated by P value. (F) Western blot shows reduction of Runx2 expression in MM.1R cells transduced with Runx2 shRNA compared with wild-type (WT) or NT control cells. (G) Six weeks after s.c. injection of NT or Runx2 k/d human MM.1R cells, serum human Ig λ light chain was measured by ELISA. Error bars represent mean ± SEM (n = 10 animals per group). Significant differences between groups are indicated by P value.

    Journal: Blood

    Article Title: Myeloma cell–derived Runx2 promotes myeloma progression in bone

    doi: 10.1182/blood-2014-12-613968

    Figure Lengend Snippet: Runx2 knockdown inhibits MM progression in vivo. (A) Expression of Runx2 in 2 Runx2 k/d 5TGM1 cell lines (Runx2 shRNA #90– and Runx2 shRNA #91–transfected 5TGM1 cells) was probed by western blot. Both Runx2 shRNA #90 and #91 decreased the expression of Runx2 compared with NT control. (B) Serum IgG2bκ was measured by ELISA 6 weeks after IV injection of NT control or Runx2 k/d 5TGM1 cells. Error bars represent mean ± SEM (n = 10 animals per group). Significant differences between groups are indicated by P value. (C) H&E-stained bone sections from mice injected IV with either NT control or Runx2 k/d 5TGM1 cells. Tumors were present in mice injected with NT cells, but not in the mice injected with Runx2 k/d cells (original magnification, ×100). Inset, Abundant myeloma cells in NT-bearing mice compared with Runx2 k/d 5TGM1 cell-injected mice. (D) Survival was significantly increased in mice injected IV with 5TGM1 Runx2 k/d cells (clone #90 and #91) compared with those injected with NT 5TGM1 cells. (E) Six weeks after intratibial injection of Runx2 k/d or NT 5TGM1 cells, levels of serum IgG2bκ were measured by ELISA. Error bars represent mean ± SEM (n = 10 animals per group). Significant differences between groups are indicated by P value. (F) Western blot shows reduction of Runx2 expression in MM.1R cells transduced with Runx2 shRNA compared with wild-type (WT) or NT control cells. (G) Six weeks after s.c. injection of NT or Runx2 k/d human MM.1R cells, serum human Ig λ light chain was measured by ELISA. Error bars represent mean ± SEM (n = 10 animals per group). Significant differences between groups are indicated by P value.

    Article Snippet: The levels of human Ig λ light chain or mouse IgG2bκ in mouse sera were measured using human Ig λ or mouse IgG2bκ ELISA kits (Bethyl Laboratories Inc), respectively.

    Techniques: Knockdown, In Vivo, Expressing, shRNA, Transfection, Western Blot, Control, Enzyme-linked Immunosorbent Assay, IV Injection, Staining, Injection, Transduction