JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB266248

Human LMAN1 knockout HEK-293T cell line

Be the first to review this product! Submit a review

|

(0 Publication)

LMAN1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
5 Images
Western blot - Human LMAN1 knockout HEK-293T cell line (AB266248)
  • WB

Lab

Western blot - Human LMAN1 knockout HEK-293T cell line (AB266248)

Lanes 1 - 4 : Merged signal (red and green). Green - ab118407 observed at 55 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.

ab118407 was shown to react with LMAN1 in wild-type HEK-293T cells in western blot with loss of signal observed in LMAN1 knockout cell line ab266248 (LMAN1 knockout cell lysate ab257505). Wild-type and LMAN1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab118407 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4° at a 1 in 200 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-LMAN1 antibody [OTI1B8] (<a href='/en-us/products/primary-antibodies/lman1-antibody-oti1b8-ab118407'>ab118407</a>) at 1/200 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

LMAN1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human LMAN1 knockout HEK-293T cell line (ab266248)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 58 kDa

Observed band size: 55 kDa

false

Western blot - Human LMAN1 knockout HEK-293T cell line (AB266248)
  • WB

Lab

Western blot - Human LMAN1 knockout HEK-293T cell line (AB266248)

Lanes 1-4 : Merged signal (red and green). Green - ab126720 observed at 55 kDa. Red - loading control ab8245 observed at 37 kDa.

ab126720 Anti-LMAN1 antibody [EPR6980] was shown to specifically react with Protein ERGIC-53 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266248 (knockout cell lysate ab257505) was used. Wild-type and Protein ERGIC-53 knockout samples were subjected to SDS-PAGE. ab126720 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 Dilution and 1 in 20000 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-LMAN1 antibody [EPR6980] (<a href='/en-us/products/primary-antibodies/lman1-antibody-epr6980-ab126720'>ab126720</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

LMAN1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human LMAN1 knockout HEK-293T cell line (ab266248)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 58 kDa

Observed band size: 55 kDa

false

Western blot - Human LMAN1 knockout HEK-293T cell line (AB266248)
  • WB

Lab

Western blot - Human LMAN1 knockout HEK-293T cell line (AB266248)

Lanes 1-4 : Merged signal (red and green). Green - ab125006 observed at 55 kDa. Red - loading control ab8245 observed at 37 kDa.

ab125006 Anti-LMAN1 antibody [EPR6979] was shown to specifically react with Protein ERGIC-53 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266248 (knockout cell lysate ab257505) was used. Wild-type and Protein ERGIC-53 knockout samples were subjected to SDS-PAGE. ab125006 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 Dilution and 1 in 20000 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-LMAN1 antibody [EPR6979] (<a href='/en-us/products/primary-antibodies/lman1-antibody-epr6979-ab125006'>ab125006</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

LMAN1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human LMAN1 knockout HEK-293T cell line (ab266248)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 58 kDa

Observed band size: 55 kDa

false

Sanger Sequencing - Human LMAN1 knockout HEK-293T cell line (AB266248)
  • Sanger seq

Unknown

Sanger Sequencing - Human LMAN1 knockout HEK-293T cell line (AB266248)

Homozygous : 1 bp insertion in exon 1

Cell Culture - Human LMAN1 knockout HEK-293T cell line (AB266248)
  • Cell Culture

Unknown

Cell Culture - Human LMAN1 knockout HEK-293T cell line (AB266248)

Representative images of LMAN1 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1

style
left-column

Complementary Products

Cell Lines & Lysates

AB266323

Human PDCD6 (ALG-2) knockout HEK-293T cell line

Advanced Validation

Western blot - Human PDCD6 (ALG-2) knockout HEK-293T cell line (AB266323)

  • 0
  • 0
Cell Lines & Lysates

AB266343

Human RAD23A (hHR23A) knockout HEK-293T cell line

Advanced Validation

Western blot - Human RAD23A (hHR23A) knockout HEK-293T cell line (AB266343)

  • 0
  • 0
Primary Antibodies

AB125006

Anti-LMAN1 antibody [EPR6979]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LMAN1 antibody [EPR6979] (AB125006)

  • 5
  • 4
Cell Lines & Lysates

AB266261

Human PCBD1 knockout HEK-293T cell line

Advanced Validation

Western blot - Human PCBD1 knockout HEK-293T cell line (AB266261)

  • 0
  • 0
style
left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
style
left-column

Product details

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

style
left-column

What's included?

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab266248-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab255449 Human wild-type HEK-293T cell line", "number":"AB266248-CMP02" }, { "size":"1 x 1000000 Cells/vial", "name":"ab266248 Human LMAN1 knockout HEK-293T cell line", "number":"AB266248-CMP01" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab266248-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab266248 Human LMAN1 knockout HEK-293T cell line", "number":"AB266248-CMP01", "productcode":"" } ] } } }
style
left-column

Properties and storage information

Gene name
LMAN1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
style
left-column

Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

style
left-column

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

LMAN1 also known as ERGIC-53 or lectin mannose-binding 1 is a protein involved in the transport and quality control of glycoproteins. It has a mass of approximately 53 kDa. LMAN1 primarily expresses in the endoplasmic reticulum-Golgi intermediate compartment (ERGIC) and is important for efficient cargo sorting and proper folding. It functions by binding to specific carbohydrate moieties on glycoproteins aiding their transport between the ER and Golgi apparatus.
Biological function summary

LMAN1 functions significantly within a protein complex known as the LMAN1-MCFD2 complex. It serves an essential role as a cargo receptor for glycoproteins particularly those with particular mannose-rich domains influencing their secretion and stability. This protein complex is integral in maintaining protein homeostasis and cellular physiology by ensuring the correct sorting and transport of glycoproteins from the ER to the Golgi.

Pathways

LMAN1 significantly participates in the secretory pathway and quality control mechanisms in the cell. It interacts with proteins such as MCFD2 to mediate the transport of specific glycoproteins like α1-antitrypsin through the secretory pathway. This pathway is important for the secretion of a variety of proteins and disruptions can lead to disorders associated with improper protein folding and trafficking.

Defects in LMAN1 can lead to bleeding disorders such as combined coagulation factor V and VIII deficiency. The protein's dysfunction impairs the transport of coagulation factors disrupting their secretion. Additionally proteins like MCFD2 work closely with LMAN1 in this context as mutations in either protein can cause a similar deficiency highlighting the significant role of LMAN1 in maintaining normal hemostatic processes.
style
left-column
style
left-column

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

style
left-column

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

style
left-column

Product protocols

style
left-column

Target data

See full target information LMAN1
style
left-column
style
left-column
style
right-column

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com