CKLF6 Antibody | Affinity Biosciences

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Western blot analysis of extracts from COS-7, using CKLF6 Antibody.

Figure 6 ATP11B interacts with PD-L1 in a CMTM6-dependent manner.

Product:	CKLF6 Antibody
Catalog:	DF3944
Description:	Rabbit polyclonal antibody to CKLF6
Application:	WB IF/ICC
Reactivity:	Human, Mouse, Monkey
Prediction:	Horse, Rabbit
Mol.Wt.:	25 KD; 20kD(Calculated).
Uniprot:	Q9NX76
RRID:	AB_2836297

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

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MS Report

HPLC Report

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:1000, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human,Mouse,Monkey

Prediction:

Horse(91%), Rabbit(100%)

Clonality:

Polyclonal

Specificity:

CKLF6 Antibody detects endogenous levels of total CKLF6.

RRID:

AB_2836297
Cite Format: Affinity Biosciences Cat# DF3944, RRID:AB_2836297.

Conjugate:

Unconjugated.

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Storage:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

Alias:

Fold/Unfold

chemokine like factor super family 6; chemokine like factor superfamily 6; chemokine like factor superfamily member 6; Chemokine-like factor superfamily member 6; CKLF like MARVEL transmembrane domain containing 6; CKLF-like MARVEL transmembrane domain-containing protein 6; CKLF6_HUMAN; CKLFSF6; Cmtm6; Da2 17; LRRGT00102; PRO2219;

Immunogens

Immunogen:

Uniprot:

Q9NX76(CKLF6_HUMAN) >>Visit HPA database.

Gene(ID):

CMTM6(54918)

Expression:

Q9NX76 CKLF6_HUMAN:

Expressed in the leukocytes, placenta and testis.

Sequence:

Q9NX76 CKLF6_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MENGAVYSPTTEEDPGPARGPRSGLAAYFFMGRLPLLRRVLKGLQLLLSLLAFICEEVVSQCTLCGGLYFFEFVSCSAFLLSLLILIVYCTPFYERVDTTKVKSSDFYITLGTGCVFLLASIIFVSTHDRTSAEIAAIVFGFIASFMFLLDFITMLYEKRQESQLRKPENTTRAEALTEPLNA

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Rabbit

100

Horse

Dog

Pig

Bovine

Sheep

Xenopus

Zebrafish

Chicken

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q9NX76 As Substrate

Q9NX76

Site	PTM Type	Source
M1	Acetylation	Uniprot
Y7	Phosphorylation	Uniprot
S8	Phosphorylation	Uniprot
Y28	Phosphorylation	Uniprot
T171	Phosphorylation	Uniprot
T178	Phosphorylation	Uniprot

Research Backgrounds

Q9NX76_CKLF6_HUMAN

Function:

Master regulator of recycling and plasma membrane expression of PD-L1/CD274, an immune inhibitory ligand critical for immune tolerance to self and antitumor immunity. Associates with both constitutive and IFNG-induced PD-L1/CD274 at recycling endosomes, where it protects PD-L1/CD274 from being targeted for lysosomal degradation, likely by preventing its STUB1-mediated ubiquitination. May stabilize PD-L1/CD274 expression on antigen presenting cells and potentiates inhibitory signaling by PDCD1/CD279, its receptor on T-cells, ultimately triggering T-cell anergy.

Subcellular Location:

Cell membrane>Multi-pass membrane protein. Early endosome membrane>Multi-pass membrane protein. Recycling endosome membrane>Multi-pass membrane protein.
Note: Co-localizes with PD-L1/CD274 in the plasma membrane and in recycling endosomes.

Tissue Specificity:

Expressed in the leukocytes, placenta and testis.

Subunit Structure:

Interacts with PD-L1/CD274 (via transmembrane domain); the interaction is direct. Interacts with CMTM4. Interacts with CD58, ARG1, ENO1 and TMPO.

Family&Domains:

Belongs to the chemokine-like factor family.

References

1). Oncolytic peptide LTX-315 induces anti-pancreatic cancer immunity by targeting the ATP11B-PD-L1 axis. Journal for ImmunoTherapy of Cancer, 2022 (PubMed: 35288467) [IF=10.9]

Application: WB Species: Human Sample: pancreatic cancer tissue

Figure 6 ATP11B interacts with PD-L1 in a CMTM6-dependent manner. (A–B) ATP11B interaction with PD-L1 in vivo. Cell lysates from KPC and SW1990 cells were separately subjected to immunoprecipitation and western blotting using the indicated antibodies. (C) Lack of binding of ATP11B to PD-L1 in vitro. FLAG-tagged ATP11B and MYC-tagged PD-L1 purified from 293 T cell lysates and subjected to immunoprecipitation after co-incubation. (D) Positive correlation of ATP11B with CMTM6. Correlation of ATP11B and CMTM6 analyzed according to the pancreatic data sets of The Cancer Genome Atlas. (E–G) Representative images (E–F) and statistical results (G) of CMTM6 and ATP11B immunohistochemical staining in pancreatic cancer tissue microarray. (H) Western blotting of ATP11B, CMTM6, and PD-L1 expression in paired clinical tissue samples. (I–M) Interaction of ATP11B with PD-L1 in pancreatic cancer in a CMTM6-dependent manner. (I−J) cell lysates from KPC and SW1990 cells were separately subjected to immunoprecipitation and western blotting using the indicated antibodies. (K) Western blotting of ATP11B expression in pancreatic cancer cell lines after CMTM6 knockdown. (L–M) Cell lysates from CMTM6-KD KPC (L) and CMTM6-KD SW1990 (M) separately subjected to immunoprecipitation and western blotting using the indicated antibodies. CMTM6, CKLF-like MARVEL transmembrane domain containing 6; PD-L1 programmed cell death ligand 1.

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CKLF6 Antibody - #DF3944