Recombinant
RabMAb

Recombinant Anti-Laminin gamma 1 antibody [EPR21199] (ab233389)

Overview

  • Product name

    Anti-Laminin gamma 1 antibody [EPR21199]
    See all Laminin gamma 1 primary antibodies
  • Description

    Rabbit monoclonal [EPR21199] to Laminin gamma 1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human Laminin gamma 1 aa 1000-1150. The exact sequence is proprietary.
    Database link: P11047

  • Positive control

    • WB: A431, NIH:OVCAR-3, HeLa, MCF7, C6, PC-12 and A549 whole cell lysates; Human fetal heart and fetal kidney tissue lysates; rat kidney, brain and spleen tissue lysates; mouse brain tissue lysate. IP: A431 whole cell lysate. IHC-P: Human pancreas tissue; mouse kidney tissue; rat stomach tissue. Flow cyt: A431 cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab233389 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 210-250 kDa (predicted molecular weight: 178 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/50.
IP 1/30.

Target

  • Function

    Binding to cells via a high affinity receptor, laminin is thought to mediate the attachment, migration and organization of cells into tissues during embryonic development by interacting with other extracellular matrix components.
  • Tissue specificity

    Found in the basement membranes (major component).
  • Sequence similarities

    Contains 11 laminin EGF-like domains.
    Contains 1 laminin IV type A domain.
    Contains 1 laminin N-terminal domain.
  • Domain

    The alpha-helical domains I and II are thought to interact with other laminin chains to form a coiled coil structure.
    Domains VI and IV are globular.
  • Cellular localization

    Secreted > extracellular space > extracellular matrix > basement membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • LAMB2 antibody
    • lamc1 antibody
    • LAMC1_HUMAN antibody
    • Laminin B2 chain antibody
    • Laminin subunit gamma-1 antibody
    • Laminin, gamma 1 (formerly LAMB2) antibody
    • Laminin-1 subunit gamma antibody
    • Laminin-10 subunit gamma antibody
    • Laminin-11 subunit gamma antibody
    • Laminin-2 subunit gamma antibody
    • Laminin-3 subunit gamma antibody
    • Laminin-4 subunit gamma antibody
    • Laminin-6 subunit gamma antibody
    • Laminin-7 subunit gamma antibody
    • Laminin-8 subunit gamma antibody
    • Laminin-9 subunit gamma antibody
    • MGC87297 antibody
    • S-LAM gamma antibody
    • S-laminin subunit gamma antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling Laminin gamma 1 with ab233389 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in basement membrane of human pancreas (PMID: 15159456) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized A431 (human epidermoid carcinoma cell line) cell line labeling Laminin gamma 1 with ab233389 at 1/50 (red) compared with Rabbit monoclonal IgG Isotype control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

  • Laminin gamma 1 was immunoprecipitated from 0.35 mg of A431 (human epidermoid carcinoma cell line) whole cell lysate with ab233389 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab233389 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

    Lane 1: A431 whole lysate 10 μg (Input).
    Lane 2: ab233389 IP in A431 whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab233389 in A431 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 15 seconds.

  • Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling Laminin gamma 1 with ab233389 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in basement membrane of rat stomach (PMID: 12571278) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Laminin gamma 1 with ab233389 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in the basement membrane of mouse kidney (PMID: 21903675) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • All lanes : Anti-Laminin gamma 1 antibody [EPR21199] (ab233389) at 1/5000 dilution

    Lane 1 : C6 (rat glial tumor glial cell line) whole cell lysate at 10 µg
    Lane 2 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 20 µg
    Lane 3 : Rat heart lysate at 10 µg
    Lane 4 : Rat kidney lysate at 10 µg
    Lane 5 : Rat brain lysate at 10 µg
    Lane 6 : Rat spleen lysate at 10 µg
    Lane 7 : Mouse brain lysate at 10 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 178 kDa
    Observed band size: 210-250 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times.

    Lane 1: 70 seconds. Lane 2: 15 seconds. Lanes 3 & 4: 10 seconds. Lanes 5 & 6: 3 minutes. Lane 7: 48 seconds.

    The expression profile observed is consistent with what has been described in the literature (PMID: 18263645, 11067943, 23857767, 28928891).

  • Lanes 1-3 & 6-7 : Anti-Laminin gamma 1 antibody [EPR21199] (ab233389) at 1/5000 dilution
    Lanes 4-5 : Anti-Laminin gamma 1 antibody [EPR21199] (ab233389) at 1/1000 dilution

    Lane 1 : A431 (human epidermoid carcinoma cell line) whole cell lysate
    Lane 2 : NIH:OVCAR-3 (human ovary adenocarcinoma cell line) whole cell lysate
    Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 4 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
    Lane 5 : A549 (human lung carcinoma cell line) whole cell lysate
    Lane 6 : Human fetal heart lysate
    Lane 7 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lanes 1-5 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lanes 6-7 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

    Predicted band size: 178 kDa
    Observed band size: 210-250 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times.

    Lanes 1, 2 & 7: 15 seconds. Lane 3: 8 seconds. Lanes 4: 37 seconds. Lanes 5 & 6: 10 seconds.

    The expression profile observed is consistent with what has been described in the literature (PMID: 18263645, 11067943, 23857767, 28928891).

References

ab233389 has not yet been referenced specifically in any publications.

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